CN104306083A - Bionic structure with passageways as well as electromagnetic force training device and electromagnetic force training method thereof - Google Patents
Bionic structure with passageways as well as electromagnetic force training device and electromagnetic force training method thereof Download PDFInfo
- Publication number
- CN104306083A CN104306083A CN201410535181.8A CN201410535181A CN104306083A CN 104306083 A CN104306083 A CN 104306083A CN 201410535181 A CN201410535181 A CN 201410535181A CN 104306083 A CN104306083 A CN 104306083A
- Authority
- CN
- China
- Prior art keywords
- cell
- passage
- biomimetic features
- pulsation
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
Abstract
The invention discloses a bionic structure with passageways as well as an electromagnetic force training device and an electromagnetic force training method thereof, and belongs to the field of the biological tissue engineering machine biomedical instruments. The electromagnetic force training device is used for training the multi-functional and multi-system three-dimensional bionic structure with the passageways in vitro; a structure main body of each passageway is in a state with two through holes in two ends, a state with two blind holes in two ends or a state with a blind hole in one end and a through hole in the other end; the passageways are intersected, parallel, collinear or antarafacial. The bionic structure is trained or subjected to pulsation training in an electric field, a magnetic field or a compound electromagnetic field, so that cells are oriented and distributed layer by layer in micro fluid passageways. The structure comprises at least one type of cells; at least one type of cells is distributed in the outer walls, inner walls or holes of the passageways. A main body of the bionic structure is a mixture of cells and natural polymer water gel, and the multiple passageways have circulation, nerve and immunity functions. The structure is a substitution for a tissue or an organ of a human body and can be used for providing a carrier for high-throughput drug screening and providing the possibility for organ transplantation.
Description
Technical field
The invention belongs to histoorgan manufacture, biological engineering and biomedical devices field, relate to a kind of biomimetic features containing passage and preparation method thereof.
Background technology
At present, organ transplantation technique is faced with immunologic rejection all the time, and donor shortage, organ distributes, the series of problems such as ethics.According to statistics, China needs the patient carrying out organ transplantation more than 1,500,000 every year, but supply and demand are than less than 1:100 [Langer R.Tissue Engineering, 2007,13 (1): 1 ~ 2].The concept that tissue engineering (Tissue Engineering) and organ manufacture (Organ Manufacturing) utilizing works and medical science combine is that this difficult problem has had brand-new solution route from tissue and organ aspect.
The biomimetic features of artificial organ or organ can be used for the repair tissue defect that implants, substitute organ function; Also can or as a kind of device outside, temporary transient substitute organ function [Cao Yilin etc., Clinical Surgery's magazine, 2007,15 (1): 40-41].At present, the histoorgan utilizing tissue engineering method to build achieves application clinically, but these application are confined to the tissue such as cartilage, epithelium more, and these organizational structuries are simple, and composition is single, size-constrained.And in the tissue engineering of complex organ manufactures, the main bugbear that people face is, while constructing function tissue and supporting structure, builds corresponding multi-functional organ substitute, and these functions mainly comprise blood circulation, nervous system and immune system.
Important organ in human body, as heart, kidney and liver etc., has complicated blood vessel, nerve, immunity and hormonal system.The control environment of cell survival is in the scope of 150 ~ 200 μm around blood vessel, otherwise freely can not cause death because of material exchange very soon; And to set up neural contact be the important channel that transplant organ exchanges with human body, this is to complex organ functionating, participates in human body regulating system and all has great significance.To sum up, how the biomimetic features containing passage of structure multisystem compound is inevitable problem during complex organ manufactures.
In addition, though the complicated conduit that the current tissue/organ biomimetic features containing passage has pipeline configuration to come in analogue body, but these pipelines are difficult to form controlled single or multiple lift cell layer structure in microcosmic point, and this will be attributed to current forming technique limitation on a microscopic scale.Therefore, we wish the effect adopting impressed field, control the attachment of cell monolayer in microfluidic channel on a microscopic scale and align.The present invention adopts the effect of extra electric field, magnetic field or composite electromagnetic field, carry out arrangement to charged particle (mainly solution particle, ordinary cells, magnetic cell and galvanic cells) to control, make cell can be arranged on duct wall according to the default number of plies, obtain the cell arrangement pattern of more analog inner catheter, reach the accurate operation to cell.
Organ manufacturing center of Tsing-Hua University (Center of Organ Manufacturing) adopts 3D printing technique and rotation combination die methods successfully to prepare the tissue or organ precursor with certain function.Utilize existing list/bis-shower nozzle Rapid Prototyping technique (also claiming 3D printing technique), [the Wang X such as simple vasoganglion, hepatic tissue and bone renovating material are successfully prepared at this center, et al.Trends in Biotechnology, 2007,25:505; Wang X.Artificial organs, 2012,36:591].Utilize rotation combination die methods [patent 201210324600.4], vascularized organ's precursor with passage has been designed at this center.But the structural limitation that these technology can be prepared at present, at simple tissue, only containing the single channel of class blood vessel, is not applied to the tissue/organ of multisystem (blood vessel, nerve and immunity) compound.In addition, existing way only can prepare general tubular structure, still can not pour into or positioning cells in tubular structure hole or on tube wall.Therefore we propose to utilize based on many shower nozzles 3D impact system and many inner cores rotation combination die methods, adopt the effect of impressed field, control the attachment of cell monolayer in microfluidic channel on a microscopic scale and align, prepare the biomimetic features more pressing close to tissue or organ, this biomimetic features is compounded with the functions such as blood circulation, nervous system, immune system and hormonal system, complex organ manufacturing technology is advanced further.
By above analysis, the histoorgan substitute pattern under prior art condition and function singleness, often only have a kind of circulation decorum (blood vessel network) of deficient maturation, not perfect in complex organ manufacture and Function.3D impact system or rotation combination die methods being combined with organ manufacturing technology is the study hotspot of medical science and engineering.
Summary of the invention
The object of this invention is to provide a kind of biomimetic features containing passage, this structure possesses the cell pipeline containing blood vessel, nerve and disposable system, makes it have the 26S Proteasome Structure and Function of actual organ.Another object of the present invention is to provide a kind of method of electromagnetic force training devices and training passage biomimetic features thereof, cell is completed in electric field, magnetic field or composite electromagnetic field and successively locates, more press close to the cell arrangement of organism internal pipeline.
Technical scheme of the present invention is as follows:
Containing a biomimetic features for passage, it is characterized in that: described biomimetic features comprises main structure body and at least one passage, and described passage is distributed in main structure body; Described main structure body is the mixture of natural polymer hydrogel and cell composition, and described cell concentration is 10-10
8individual/mL, this cell is at least one in embryonic stem cell, adult stem cell, adult cell, cancerous cell and induced multi-potent stem cells; Described passage is single channel or branched bottom, and passage is the one or more combination in two ends through hole, two ends blind hole and blind hole one end, one end through-hole structure; Position relationship between passage is crossing, parallel, conllinear or antarafacial; Be distributed with cell in described channel outer wall, vias inner walls or access opening, cell is at least two kinds in nervous system seed cell, vascular system seed cell and immune system seed cell.
In technique scheme, the cross section of described passage is circular, ellipse, polygon or irregular geometric figures, and cross sectional area is 100 μm
2-1cm
2.Described a kind of biomimetic features containing passage, is characterized in that: described nervous system seed cell is at least one in neuron and neurogliocyte; Described blood vessel seed cell is at least one in endotheliocyte, smooth muscle cell and fat stem cell; Described immune system seed cell is at least one in lymphocyte and inherent immunity cell.
Natural polymer hydrogel of the present invention is at least one in sodium alginate, collagen, matrigel, dextrose, chitosan, gelatin and Fibrinogen, and the mass body volume concentrations of this hydrogel is 0.1 ~ 20%.At least one in cell cryopreservation agent, cell growth factor, medicine, anticoagulant and magnetic nanoparticle is compounded with in described natural polymer hydrogel; Described cell cryopreservation agent is at least one in dimethyl sulfoxide, glycerol and dextrose; Described cell growth factor is at least one in VEGF, basic fibroblast growth factor, hepatocyte growth factor, human blood platelets derived growth factor and transforming growth factor; Described medicine is at least one in antitumor drug and viral vaccine; Described anticoagulant is at least one in heparin and paclitaxel; Described magnetic nanoparticle is at least one in ferrite particle, metal mold granule and nitrided iron granule.
The electromagnetic force training devices of a kind of biomimetic features containing passage provided by the invention, is characterized in that: described device comprises electric field generation system, magnetic field generation system, sample stage and fixed platform; Described magnetic field generation system and sample stage are installed in fixed platform; Described electric field generation system is containing positive pole and negative pole; Described sample stage is provided with guide rail, and described positive pole and negative pole are arranged on guide rail respectively by slide block; Described sample stage is positioned at magnetic field generation system; Described magnetic field generation system divides and comprises support and rotatable annulus, and described rotatable annulus is arranged in the hollow structure of support, and on rotatable annulus, radial symmetric is placed with dismountable S pole and N pole.
Also pulsation culture systems can be comprised in device of the present invention; Described pulsation culture systems is installed in fixed platform; Described pulsation culture systems comprises pulsation system motor, pulsation system guide rail-slide block mechanism, culture fluid supply syringe, check valve, mozzle and culture jar; Described pulsation system motor is connected with pulsation system guide rail-slide block mechanism by crank; Described pulsation system guide rail-slide block mechanism supplies syringe with culture fluid and is connected.
In device of the present invention, described positive pole and negative pole are the one in metal plate, plain conductor or metal probe.Described S pole and the N one very in permanent magnet or electric magnet.
Present invention also offers a kind of electric field of said apparatus that utilizes to train the method for biomimetic features, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) start the electric field generation system of electromagnetism training devices, remove N level and the S level of magnetic field generation system; The biomimetic features containing passage is made to be arranged in electric field; Electric current, for being greater than 0, is less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly;
C) complete electric field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Present invention also offers a kind of magnetic field of said apparatus that utilizes to train the method for biomimetic features, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) close the electric field generation system of electromagnetism training devices, start magnetic field generation system, S pole and N pole are rotated with rotatable annulus, controlling magnetic induction for being greater than 0, being less than or equal to 5T;
C) complete magnetic field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Present invention also offers a kind of electromagnetic complex field of said apparatus that utilizes to train the method for biomimetic features, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) start electric field generation system and the magnetic field generation system of electromagnetism training devices simultaneously, make the biomimetic features containing passage be arranged in Electric and magnetic fields; Controlling electric current for being greater than 0, being less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly; Controlling magnetic induction for being greater than 0, being less than or equal to 5T;
C), after completing the training of Electric and magnetic fields simultaneously to the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Train on the basis of the method for biomimetic features at above-mentioned electric field, magnetic field and electromagnetic complex field, the invention provides and pulsation culture systems is applied training method wherein simultaneously, it is characterized in that: start pulsation culture systems, described pulsation culture systems comprises pulsation system motor, pulsation system guide rail-slide block mechanism, culture fluid supply syringe, check valve, mozzle and culture jar; Described pulsation system motor is connected with pulsation system guide rail-slide block mechanism by crank; Described guide rail-slide block mechanism and culture fluid supply syringe and are connected; By mozzle, described biomimetic features is connected with pulsation culture systems; Culture fluid is made to carry out one-way flow between mozzle and the biomimetic features containing passage; Flow rate of liquid, for being greater than 0, is less than 30cm/s, the effect of-electric field that carries out pulsing, pulsation-magnetic field or pulsation-electromagnetic complex field; After completing the training to described biomimetic features, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
The present invention compared with prior art, has the technique effect of following advantage and salience:
1. the present invention is by existing three-dimensional technology as based on 3D impact system and rotation combination die methods, by cell and the further compound of high-molecular gel mixture, prepares passage, multisystem, multi-functional complex organization device biomimetic features.Vascular system, nervous system and immune system should be enumerated containing the biomimetic features of passage, structurally and functionally simulate the organ state of real multisystem in body dramatically.
2. a kind of electromagnetic force training devices of the present invention, combine the effects such as electric field, magnetic field and pulsation cultivation, external training and mechanics cultivation are carried out to biomimetic features, cell can be made successively to locate under the effect of Composite Field, be arranged in pipeline configuration periphery, press close to the cell morphology of actual organ's conduit, structure formed thereby has good performance on morphosis, in immune physiology.
3. the using method of a kind of electromagnetic force training devices of the present invention, can be used alone electric field, is used alone magnetic field or electromagnetic field is met use simultaneously; In addition, the introduction of culture systems of pulsing too increases the diversification of apparatus function.
Accompanying drawing explanation
Fig. 1 is the bionical organizational structure schematic diagram containing passage.
Fig. 2 a ~ 2i is the view of several passage cell, wherein, Fig. 2 a is the schematic diagram that a kind of cell is positioned at channel outer wall, Fig. 2 b is the schematic diagram that a kind of cell is positioned at vias inner walls, Fig. 2 c is the schematic diagram that a kind of cell is positioned at access opening, Fig. 2 d is the schematic diagram that two kinds of cells are positioned at channel outer wall, Fig. 2 e is the schematic diagram that two kinds of cells are positioned at vias inner walls, Fig. 2 f is the schematic diagram that two kinds of cells are positioned at access opening, Fig. 2 g is the schematic diagram that a kind of cell is positioned at that the another kind of cell of channel outer wall is positioned at access opening, Fig. 2 h is the schematic diagram that a kind of cell is positioned at that the another kind of cell of vias inner walls is positioned at access opening, Fig. 2 i is the schematic diagram that a kind of cell is positioned at that the another kind of cell of channel outer wall is positioned at vias inner walls.
Fig. 3 adopts many shower nozzles 3D impact system to be shaped containing the biomimetic features schematic diagram of passage.
Fig. 4 a and Fig. 4 b is respectively schematic diagram and the explosive view of many inner cores rotation combination die methods formed channel biomimetic features.
Fig. 5 a, 5b and 5c are respectively fusiform dual pathways biomimetic features schematic diagram, block cross aisle biomimetic features schematic diagram and block parallel channels biomimetic features schematic diagram.
Fig. 6 is electromagnetism training devices schematic diagram.
Fig. 7 a is magnetic field generation system exploded perspective view, and Fig. 7 b is pulsation culture systems schematic diagram.
Fig. 8 a is containing the biomimetic features of passage between metal plate electrode, Fig. 8 b be the biomimetic features of passage between plain conductor or metal probe, Fig. 8 c is the motion schematic diagram of charged particle in electric field (E).
Fig. 9 a is that Fig. 9 b is the motion schematic diagram of magnetic particle in magnetic field (B) (dotted line represents magnetic field rotation direction) containing the biomimetic features of passage between magnetic field S, N pole.
Figure 10 a is that Figure 10 b is the motion schematic diagram of charged particle in composite electromagnetic field (dotted line represents magnetic field rotation direction) containing the biomimetic features of passage between electric field positive and negative electrode and magnetic field S, N pole.
In figure: 101-main structure body; 102-is positioned at the cell of main body; The passage of 103-two ends through hole; The passage of blind hole one end, 104-one end through hole; The passage of 105-two ends blind hole; 106-is containing branched bottom; 107-is positioned at the cell of access opening; 108-is positioned at the cell of channel outer wall; 109-is positioned at the cell of vias inner walls; 301-3D printing head assembly; The main structure body that 302-3D prints; The passage of the circular cross-section that 303-3D prints; The passage of the irregular section that 304-3D prints; 401-assembling die base; 402-assembling die branch inner core; 403-assembling die mold; 404-assembling die base jack; 501-is containing multichannel biomimetic features main body; 502-is containing the passage of multichannel biomimetic features; 601-electric field generation system (positive pole and negative pole); 602-support; 603-sample stage; 604-motor drives; 605-fixed platform; 606-N pole; 607-S pole; The rotatable annulus of 608-; 701-is pulsed culture systems; 702-pulsation system motor; 703-pulsation system guide rail-slide block mechanism; 704-culture fluid supply syringe; 705-check valve; 706-mozzle; 707-fusiform passage biomimetic features; 708-culture jar.
Detailed description of the invention
Below in conjunction with drawings and Examples, the present invention is further described.
As shown in Figure 1, a kind of biomimetic features containing passage, comprise main structure body 101 and at least one passage, described passage is distributed in main structure body 101; Described main structure body is the mixture of natural polymer hydrogel and cell composition, and described cell concentration is 10-10
8individual/mL, this cell is at least one in embryonic stem cell, adult stem cell, adult cell, cancerous cell and induced multi-potent stem cells; Described passage is single channel or branched bottom 105, and passage is the one or more combination in two ends through hole 103, two ends blind hole 106 and blind hole one end, one end through-hole structure 104; Position relationship between passage is crossing, parallel, conllinear or antarafacial; Be distributed with cell in described channel outer wall, vias inner walls or access opening, cell is at least two kinds in nervous system seed cell, vascular system seed cell and immune system seed cell.The cross section of described passage is circular, ellipse, polygon or irregular geometric figures, and cross sectional area is 100 μm
2-1cm
2.Described nervous system seed cell is at least one in neuron and neurogliocyte; Described blood vessel seed cell is at least one in endotheliocyte, smooth muscle cell and fat stem cell; Described immune system seed cell is at least one in lymphocyte and inherent immunity cell.
Fig. 2 a ~ 2i is the view of several passage cell, wherein, Fig. 2 a is the schematic diagram that a kind of cell is positioned at channel outer wall, Fig. 2 b is the schematic diagram that a kind of cell is positioned at vias inner walls, Fig. 2 c is the schematic diagram that a kind of cell is positioned at access opening, Fig. 2 d is the schematic diagram that two kinds of cells are positioned at channel outer wall, Fig. 2 e is the schematic diagram that two kinds of cells are positioned at vias inner walls, Fig. 2 f is the schematic diagram that two kinds of cells are positioned at access opening, Fig. 2 g is the schematic diagram that a kind of cell is positioned at that the another kind of cell of channel outer wall is positioned at access opening, Fig. 2 h is the schematic diagram that a kind of cell is positioned at that the another kind of cell of vias inner walls is positioned at access opening, Fig. 2 i is the schematic diagram that a kind of cell is positioned at that the another kind of cell of channel outer wall is positioned at vias inner walls.As shown in Fig. 3, Fig. 4 a and Fig. 4 b, it is the preparation method of biomimetic features.Fig. 3 adopts many shower nozzles 3D impact system to be shaped containing the biomimetic features schematic diagram of passage.Fig. 4 a and Fig. 4 b is respectively schematic diagram and the explosive view of many inner cores rotation combination die methods formed channel biomimetic features.
Fig. 5 a ~ 5c is several containing multichannel biomimetic features schematic diagram.Fig. 5 a, 5b and 5c are respectively fusiform dual pathways biomimetic features schematic diagram, block cross aisle biomimetic features schematic diagram and block parallel channels biomimetic features schematic diagram.Described natural polymer hydrogel is at least one in sodium alginate, collagen, matrigel, dextrose, chitosan, gelatin and Fibrinogen, and the mass body volume concentrations of this hydrogel is 0.1 ~ 20%.At least one in cell cryopreservation agent, cell growth factor, medicine, anticoagulant and magnetic nanoparticle is compounded with in described natural polymer hydrogel; Described cell cryopreservation agent is at least one in dimethyl sulfoxide, glycerol and dextrose; Described cell growth factor is at least one in VEGF, basic fibroblast growth factor, hepatocyte growth factor, human blood platelets derived growth factor and transforming growth factor; Described medicine is at least one in antitumor drug and viral vaccine; Described anticoagulant is at least one in heparin and paclitaxel; Described magnetic nanoparticle is at least one in ferrite particle, metal mold granule and nitrided iron granule.
As shown in Figure 6, a kind of electromagnetism training devices, comprises electric field generation system 601, magnetic field generation system, sample stage 603 and fixed platform 605; Described magnetic field generation system and sample stage 603 are installed in fixed platform 605; Described electric field system 601 system is containing positive pole and negative pole; Described sample stage 603 is provided with guide rail, and described positive pole and negative pole are arranged on guide rail respectively by slide block; Described sample stage 603 is positioned at magnetic field generation system; Described magnetic field generation system divides and comprises support 602 and rotatable annulus 608, and described rotatable annulus 608 is arranged in the hollow structure of support 602, and on rotatable annulus 608, radial symmetric is placed with dismountable S pole 606 and N pole 607, refers to Fig. 7 a.Described device also can comprise pulsation culture systems 701; Described pulsation culture systems is installed in fixed platform 605; Described pulsation culture systems 701 comprises pulsation system motor 702, pulsation system guide rail-slide block mechanism 703, culture fluid supply syringe 704, check valve 705, mozzle 706 and culture jar 708; Described pulsation system motor 702 is connected with pulsation system guide rail-slide block mechanism 703 by crank; Described pulsation system guide rail-slide block mechanism 703 supplies syringe 704 with culture fluid and is connected, and refers to Fig. 7 b.Described positive pole and negative pole are the one in metal plate, plain conductor or metal probe.Described S pole and the N one very in permanent magnet or electric magnet.
As shown in Figure 8 a, containing the biomimetic features of passage between metal plate electrode, Fig. 8 b is that Fig. 8 c is the motion schematic diagram of charged particle in electric field (E) containing the biomimetic features of passage between plain conductor or metal probe.By to being placed in the method that described device electric field trains containing the biomimetic features of passage, the present invention comprises the steps: that the biomimetic features containing passage that a) will prepare immerses the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices; B) start the electric field generation system of electromagnetism training devices, remove N level and the S level of magnetic field generation system; The biomimetic features containing passage is made to be arranged in electric field; Electric current, for being greater than 0, is less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly; C) complete electric field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
As illustrated in fig. 9, containing the biomimetic features of passage between magnetic field S, N pole, Fig. 9 b is the motion schematic diagram of magnetic particle in magnetic field (B) (dotted line represents magnetic field rotation direction).The present invention comprises the steps: a) biomimetic features containing passage prepared to be immersed the cell culture fluid containing suspension cell by being placed in the method for training in described device magnetic field to the biomimetic features containing passage, and is placed on the sample stage of electromagnetic force training devices; B) close the electric field generation system of electromagnetism training devices, start magnetic field generation system, S pole and N pole are rotated with rotatable annulus, controlling magnetic induction for being greater than 0, being less than or equal to 5T; C) complete magnetic field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Figure 10 a is that Figure 10 b is the motion schematic diagram of charged particle in composite electromagnetic field (dotted line represents magnetic field rotation direction) containing the biomimetic features of passage between electric field positive and negative electrode and magnetic field S, N pole.By to being placed in the method that described device electromagnetic complex field trains containing the biomimetic features of passage, the present invention comprises the steps: that the biomimetic features containing passage that a) will prepare immerses the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices; B) start electric field generation system and the magnetic field generation system of electromagnetism training devices simultaneously, make the biomimetic features containing passage be arranged in Electric and magnetic fields; Controlling electric current for being greater than 0, being less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly; Controlling magnetic induction for being greater than 0, being less than or equal to 5T; C), after completing the training of Electric and magnetic fields simultaneously to the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Train on the basis of the method for biomimetic features at above-mentioned electric field, magnetic field and electromagnetic complex field, pulsation culture systems is also applied training method wherein by the present invention simultaneously, its step comprises above-mentioned steps, comprise introducing and the use of pulsation culture systems in addition: start pulsation culture systems 701, described pulsation culture systems 701 comprises pulsation system motor 702, pulsation system guide rail-slide block mechanism 703, culture fluid supply syringe 704, check valve 705, mozzle 706 and culture jar 708; Described pulsation system motor 702 is connected with pulsation system guide rail-slide block mechanism 703 by crank; Described guide rail-slide block mechanism 703 and culture fluid supply syringe 704 and are connected; By mozzle 706, described biomimetic features is connected with pulsation culture systems; Culture fluid is made to carry out one-way flow between mozzle 706 and the biomimetic features containing passage; Flow rate of liquid, for being greater than 0, is less than 30cm/s, the effect of-electric field that carries out pulsing, pulsation-magnetic field or pulsation-electromagnetic complex field; After completing the training to described biomimetic features, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Enumerate several specific embodiment below, to understand the present invention further.
Embodiment 1: utilize many shower nozzles 3D impact system to prepare a kind of containing blood vessel and the twin-channel imitative cardiac structure of nervous system, and this imitative cardiac structure is trained in the electric field.
A) threedimensional model containing multichannel bionic heart structure described in Computer Design is utilized;
B) using myocardial cell as host cell, mix with the sodium alginate soln configured, Sodium Alginate Hydrogel Films solution quality volumetric concentration is 0.5%; Afterwards this mixture is loaded in a nozzle component of many shower nozzles 3D printing device;
C) using vascular endothelial cell and neuron as passage cell; Vascular endothelial cell is mixed with sodium alginate soln, loads in second nozzle component of 3D printing device; Neuron is mixed with cell culture fluid (DMEM), loads in the 3rd nozzle component of 3D printing device;
D) the different nozzle components of many shower nozzles 3D printing device are controlled by computer, the shaping of the myocardial cell/sodium alginate of control subject structure; The channel formation of synchronous or asynchronous controlling vascular endothelial cell/sodium alginate, obtains vascular system passage; Synchronous or asynchronous controlling neuron/DMEM, is located with in access opening, obtains nervous system passage; And adopting CaCl2 to make, sodium alginate soln is crosslinked obtains Sodium Alginate Hydrogel Films, successively piles up, obtains described containing multichannel twin-channel imitative cardiac structure;
E) by sample stage as electromagnetism training devices of the imitative cardiac structure for preparing;
F) positive pole of electromagnetism training devices and the metal plate of negative pole imitated the both sides of cardiac structure but do not contact;
G) just the electric field generation device of electromagnetism training devices is energized, and is direct current 50V by control voltage size;
H) complete electric field to after the training of imitative cardiac structure, proceeded In vitro culture.
Embodiment 2: utilize many shower nozzles 3D impact system to prepare a kind of containing the three-channel imitative Renal Structure of blood vessel, nerve and immune system, and this imitative Renal Structure is trained in the electric field.
A) threedimensional model containing multichannel bionic heart structure described in Computer Design is utilized;
B) using embryonic kidney cells as host cell, mix with the fibrinogen solution configured, fibrinogen solution mass body volume concentrations is 3%; Afterwards this mixture is loaded in a nozzle component of many shower nozzles 3D printing device;
C) using vascular endothelial cell, smooth muscle cell, neuron and T lymphocyte as passage cell; Vascular endothelial cell and smooth muscle cell are mixed with sodium alginate soln, loads in second nozzle component of 3D printing device; Neuron is mixed with sodium alginate soln, loads in the 3rd nozzle component of 3D printing device; T lymphocyte is mixed with sodium alginate soln, loads in the 4th nozzle component of 3D printing device;
D) controlled the different nozzle components of many shower nozzles 3D printing device by computer, the embryonic kidney cells/fibrinogenic shaping of control subject structure, and adopt thrombin to make fibrinogen gel; The channel formation of synchronous or asynchronous controlling capillary endothelial cell and smooth muscle cell/sodium alginate, obtains vascular system passage; The channel formation of synchronous or asynchronous controlling neuron/sodium alginate, obtains nervous system passage; The channel formation of synchronous or asynchronous controlling T lymphocyte/sodium alginate, obtains immune system passage; Adopt CaCl
2make sodium alginate soln gelation obtain Sodium Alginate Hydrogel Films, successively pile up, obtain described containing multichannel three-channel imitative Renal Structure.
E) by sample stage as electromagnetism training devices of the imitative Renal Structure for preparing;
F) positive pole of electromagnetism training devices is connected with imitative Renal Structure with the metal probe of negative pole, makes imitative Renal Structure become a circuit part;
G) just the electric field generation device of electromagnetism training devices is energized, and is direct current 50mA by control size of current;
H) complete electric field to after the training of imitative Renal Structure, proceeded In vitro culture.
Embodiment 3: utilize many shower nozzles 3D impact system to prepare a kind of imitative liver structure containing one-in-and-one-out blood vessel, nerve, bile duct and immune system Five-channel, and this imitative liver structure is trained in magnetic field.
A) threedimensional model containing multichannel bionic heart structure described in Computer Design is utilized;
B) using fat stem cell and hepatocyte as host cell, mix with the gelatin configured, gelatin/fibrinogen solution, gelatin, gelatin/fibrinogen solution mass body volume concentrations is for being respectively 20%, 10%; Afterwards this mixture is loaded in a nozzle component of many shower nozzles 3D printing device;
C) using vascular endothelial cell and bone-marrow-derived lymphocyte as passage cell (these two kinds of cells with Fe
3o
4magnetic nanoparticle mixes); Vascular endothelial cell is mixed with collagen solution, loads in second nozzle component of 3D printing device; Bone-marrow-derived lymphocyte is mixed with collagen solution, loads in the 3rd nozzle component of 3D printing device;
D) controlled the different nozzle components of many shower nozzles 3D printing device by computer, control subject structural fatty stem cell/gelatin and hepatocyte/gelatin/fibrinogenic shaping, and adopt thrombin that Fibrinogen is polymerized; The channel formation of synchronous or asynchronous controlling vascular endothelial cell/collagen, obtains two blood vessel accesses; The channel formation of synchronous or asynchronous controlling bone-marrow-derived lymphocyte/collagen, obtains immune system passage; Adopt glutaraldehyde solution to make collagen cross-linking be hydrogel, successively pile up, obtain described containing multichannel three-channel imitative liver structure.
E) by sample stage as electromagnetism training devices of the magnetic imitative liver structure for preparing;
F) by the rotation of drive and control of electric machine magnetic field generation system, magnetic field size and action time, magnetic induction line is made to pass magnetic imitative liver structure;
G) complete magnetic field to after the training of the raw structure of imitative liver, proceeded In vitro culture.
Embodiment 4: utilize many inner cores rotation combination die methods to prepare a kind of containing the two blood vessel of one-in-and-one-out and the three-channel imitative pancreas structure of nervous system, and this bionical pancreatic tissue is trained in composite electromagnetic field.
A) using beta Cell of islet as host cell, mix with the fibrinogen solution configured, in this mixture, fibrinogen solution mass body volume concentrations is 5%;
B) using fat stem cell and Schwann cell as passage cell, mix with cell culture medium (DMEM);
C) three mold drop inner cores are inserted in the array hole of mold base, backform tool is enclosed within mold base, and make mold drop inner core through the macropore of the center upper portion of backform tool;
D) step beta Cell of islet/fibrinogen mixture is filled in the inner chamber of backform tool by macropore, make mold base and backform tool produce relative rotation simultaneously, rotary speed is 30r/min, mold base and mold drop inner core geo-stationary, make the mixture of perfusion form half spindle profile, in rotary course, adopt thrombin to make conversion of fibrinogen be hydrogel;
E) remove backform tool and mold drop inner core successively, formed containing triple channel biomimetic features precursor;
F) fat stem cell/DMEM is filled in the one-in-and-one-out twin flue of triple channel biomimetic features precursor, forms vascular system passage; Schwann cell/DMEM is filled in another passage of passage biomimetic features precursor, forms nervous system passage; Finally, three-channel imitative pancreas structure is obtained.
G) by sample stage as electromagnetism training devices of the imitative pancreas structure for preparing;
H) by the positive pole of electromagnetism training devices and the metal probe of negative pole and imitative pancreas anatomical connectivity, imitative pancreas structure is made to become a circuit part;
I) just the electric field generation device of electromagnetism training devices is energized, and by control size of current, direction and action time; Electric field line is now through imitative pancreas structure;
J) by the rotation of drive and control of electric machine magnetic field generation system, magnetic field size and action time, the imitative pancreas structure that magnetic induction line is also passed;
K) complete compound field magnetic field to after the training of imitative pancreas structure, proceeded In vitro culture.
Embodiment 5: utilize many inner cores rotation combination die methods to prepare a kind of liver structure containing blood vessel, bile duct, nerve and immune system four-way, and this liver structure is trained in composite electromagnetic field.
A) do hepatocyte and fat as host cell, mix with the sodium alginate/glutin solution configured, fibrinogen solution, in this mixture, sodium alginate, Fibrinogen mass body volume concentrations are 5%; Add the DMSO of mass body volume concentrations 3% in the mixture;
B) using endotheliocyte, Schwann cell and neutrophilic granulocyte as passage cell; Endotheliocyte and Schwann cell are mixed with cell culture medium (DMEM); Neutrophilic granulocyte is mixed with sodium alginate soln;
C) four mold drop inner cores are inserted in the array hole of mold base, backform tool is enclosed within mold base, and make mold drop inner core through the macropore of the center upper portion of backform tool;
D) Fibrinogen/DMSO solution of hepatocyte and fat stem cell, sodium alginate/glutin/DMSO solution are filled in the inner chamber of backform tool by macropore, make mold base and backform tool produce relative rotation simultaneously, rotary speed is 30r/min, mold base and mold drop inner core geo-stationary, make the mixture of perfusion form half spindle profile, in rotary course, adopt CaCl
2sodium alginate is made to change hydrogel into;
E) remove backform tool and mold drop inner core successively, formed containing four-way biomimetic features precursor;
F) fat stem cell/heparin/EGF/PBS suspension is filled in first passage of four-way biomimetic features precursor, forms vascular system passage; Schwann cell PBS suspension is filled in second passage of four-way biomimetic features precursor, forms nervous system passage; Neutrophilic granulocyte/PBS suspension is filled in the 3rd passage of four-way biomimetic features precursor, and adopts CaCl
2crosslinked, form immune system passage; Finally, half spindle liver structure of four-way is obtained.By two and half spindle liver precursor sodium alginate/glutin/DMSO solution adheres together, outer spray one deck synthesis macromolecule polyurethane solution, forms a spindle liver precursor;
G) by sample stage as electromagnetism training devices of the spindle liver precursor for preparing;
H) positive pole of electromagnetism training devices and the metal plate of negative pole be placed in the both sides of spindle liver precursor but do not contact;
I) just the electric field generation device of electromagnetism training devices is energized, and is direct current 20V by control voltage, and electric field line is now through spindle liver front body structure;
J) by the rotation of drive and control of electric machine magnetic field generation system, magnetic field size and action time, the spindle liver front body structure that magnetic induction line is also passed;
K) complete compound field magnetic field to after the training of spindle liver precursor, put it in liquid nitrogen and preserve, clinical when needing recovery transplant.
Embodiment 6: utilize many inner cores rotation combination die methods to prepare a kind of cardiac structure containing blood vessel, nerve and immune system four-way, and this cardiac structure is trained in the electric field.
A) using myocardial cell as host cell, mix with the fibrinogen solution configured, in this mixture, fibrinogen solution mass body volume concentrations is 5%; Add the dextrose of mass body volume concentrations 3% in the mixture;
B) using fat stem cell, smooth muscle cell, neuron and and lymphocyte as passage cell, mix with cell culture medium (DMEM) respectively;
C) four mold drop inner cores are inserted in the array hole of mold base, backform tool is enclosed within mold base, and make mold drop inner core through the macropore of the center upper portion of backform tool;
D) cardiac muscle/Fibrinogen/dextrose solution mixture is filled in the inner chamber of backform tool by macropore, make mold base and backform tool produce relative rotation simultaneously, rotary speed is 30r/min, mold base and mold drop inner core geo-stationary, make the mixture of perfusion form half spindle profile, in rotary course, adopt thrombin to make conversion of fibrinogen be hydrogel;
E) remove backform tool and mold drop inner core successively, formed containing four-way biomimetic features precursor;
F) fat stem cell/EGF/b-FGF/ paclitaxel/DMEM is filled in first passage of four-way biomimetic features precursor, forms vascular system passage; Fat stem cell and smooth muscle cell/DMEM are filled in second passage of four-way biomimetic features precursor, form vascular system passage; Neuron/DMEM is filled in the 3rd passage of four-way biomimetic features precursor, forms nervous system passage; Lymphocyte/DMEM is filled in the 4th passage of four-way biomimetic features precursor, forms immune system lymphatic channels; Finally, the imitative cardiac structure of four-way is obtained.
G) by sample stage as electromagnetism training devices of the imitative cardiac structure for preparing;
H) positive pole of electromagnetism training devices is connected with imitative cardiac structure with the plain conductor of negative pole, makes cardiac structure become a circuit part;
I) just the electric field generation device of electromagnetism training devices is energized, and replaces 30mA by control size of current DC-AC;
J) complete electric field to after the training of cardiac structure, proceeded In vitro culture.
Embodiment 7: utilize multichannel detachable assembling die legal system standby a kind of containing the multichannel artificial skin structure of blood vessel, nerve and immune system, and this skin texture is trained at elect magnetic field.
A) using fibroblast as host cell, mix with the sodium alginate soln configured, in this mixture, sodium alginate soln mass body volume concentrations is 5%;
B) using fibroblast, fat stem cell as host cell, neuron and and lymphocyte as passage cell, horn cell, epithelial cell, as compound cells, mix respectively at cell culture medium (DMEM);
C) multichannel collapsible mold is inserted in the array hole of mold base, outer mold is enclosed within mold base;
D) fibroblast/sodium alginate soln mixture is filled in the inner chamber of backform tool, makes the mixture of perfusion form square, adopt CaCl
2sodium alginate is made to change hydrogel into;
E) remove outer mold and mold drop inner core successively, formed containing multi path artificial skin biomimetic features precursor;
F) fat stem cell/DMEM is filled in first passage of four-way biomimetic features precursor, forms vascular system passage; Fat stem cell and smooth muscle cell/DMEM are filled in second passage of four-way biomimetic features precursor, form vascular system passage; Neuron/DMEM is filled in the 3rd passage of four-way biomimetic features precursor, forms nervous system passage; Lymphocyte/DMEM is filled in the 4th passage of four-way biomimetic features precursor, forms immune system lymphatic channels; Finally, the artificial skin structure of four-way is obtained.
G) by sample stage as electromagnetism training devices of the artificial skin structure for preparing;
H) by the positive pole of electromagnetism training devices and the plain conductor of negative pole and artificial skin anatomical connectivity, artificial skin structure is made to become a circuit part;
I) just the electric field generation device of electromagnetism training devices is energized, and is exchange 20mA by control size of current;
J) complete electric field to after the training of artificial skin structure, proceeded In vitro culture.
Embodiment 8: utilize many inner cores rotation combination die methods to prepare a kind of containing the three-channel udder texture of blood vessel, nerve and immune system, and this udder texture is trained in composite electromagnetic field.
A) using fat stem cell as host cell, mix with the sodium alginate soln configured, in this mixture, sodium alginate soln mass body volume concentrations is 1%;
B) using endotheliocyte, Schwann cell and neutrophilic granulocyte as passage cell; Endotheliocyte and Schwann cell are mixed with cell culture medium (DMEM); Neutrophilic granulocyte is mixed with sodium alginate soln;
C) three mold drop inner cores are inserted in the array hole of mold base, backform tool is enclosed within mold base, and make mold drop inner core through the macropore of the center upper portion of backform tool;
D) fat stem cell/sodium alginate/DMSO solution mixture is filled in the inner chamber of backform tool by macropore, make mold base and backform tool produce relative rotation simultaneously, rotary speed is 30r/min, mold base and mold drop inner core geo-stationary, make the mixture of perfusion form half spindle profile, in rotary course, adopt CaCl
2sodium alginate is made to change hydrogel into;
E) remove backform tool and mold drop inner core successively, formed containing triple channel biomimetic features precursor;
F) endotheliocyte/DMEM is filled in first passage of triple channel biomimetic features precursor, forms vascular system passage; Schwann cell/DMEM is filled in second passage of triple channel biomimetic features precursor, forms nervous system passage; Neutrophilic granulocyte/sodium alginate soln is filled in the 3rd passage of triple channel biomimetic features precursor, and adopts CaCl
2crosslinked, form immune system passage; Finally, three-channel half spindle udder texture is obtained.By two and half spindle breast precursors extra large Fibrinogen/fat stem cell solution adheres together, outer spray one deck synthesis polyphosphazene polymer lactic acid and polyglycolic acid copolymers (PLGA) solution, form a spindle breast precursor;
G) by sample stage as electromagnetism training devices of the spindle breast precursor for preparing;
H) positive pole of electromagnetism training devices and the metal plate of negative pole be placed in the both sides of spindle breast precursor but do not contact;
I) just the electric field generation device of electromagnetism training devices is energized, and by control size of current, direction and action time; Electric field line is now through spindle breast front body structure;
J) by the rotation of drive and control of electric machine magnetic field generation system, magnetic field size and action time, the spindle breast front body structure that magnetic induction line is also passed;
K) complete compound field magnetic field to after the training of spindle breast precursor, put it in liquid nitrogen and preserve, clinical when needing recovery transplant.
Embodiment 9: utilize assembling die legal system to contain for a kind of the lobe of the lung structure that two enter scene 2 blood vessel, nerve and immune system four-way, and this lobe of the lung structure is trained in magnetic field.
A) utilize described in Computer Design containing the threedimensional model of the bionic lung impeller structure of four-way and assembling die;
B) using fat stem cell and pulmonary epithelial cells as host cell, mix with the gelatin solution configured, gelatin solution mass body volume concentrations is for being respectively 10%, wherein add the cell cryopreservation agent glycerol that mass fraction is 3%, afterwards this mixture is expelled to the relevant position of assembling die, then 1% collagen solution injected or be poured on corresponding cellular layer, within 37 DEG C static 30 minutes, making cell gelatin and collagen layer solidification, or adopt glutaraldehyde solution to make collagen change gel into;
C) using vascular endothelial cell and bone-marrow-derived lymphocyte as passage cell (these two kinds of cells with Fe
3o
4magnetic nanoparticle mixes); Vascular endothelial cell is mixed with fibrinogen solution, is expelled to two and enters in the vascular system of scene 2; Bone-marrow-derived lymphocyte is mixed with fibrinogen solution, is expelled in a lateral system; By schwann cell and fibrinogen solution hybrid injection in another branch's nervous system;
D) adopt 0.1% thrombin solution to make Fibrinogen be hydrogel, obtain the lobe of the lung structure of four-way.
E) the multichannel sample containing cell cryopreservation agent prepared is placed on sample stage, the vascular system of multichannel sample is connected with impulsive motion bioreactor;
F) flow controlling culture fluid in impulsive motion bioreactor is 30cm/s, and action time, 3h, made wherein cell formative tissue structure;
G) complete impulsive motion bioreactor to after the training of the mechanics of lobe of the lung structure, place it in culture fluid and continue to cultivate, or be placed on low temperature (as in liquid nitrogen) and preserve for a long time, or be directly used in organ transplantation.
Embodiment 10: utilize many shower nozzles 3D impact system to prepare a kind of containing blood vessel and the twin-channel imitative liver tumor structure of nervous system, and this imitative liver tumor structure is carried out in the electric field pulsation cultivation training.
A) threedimensional model containing multichannel bionical liver tumor structure described in Computer Design is utilized;
B) using hepatoma carcinoma cell as host cell, mix with the sodium alginate soln configured, Sodium Alginate Hydrogel Films solution quality volumetric concentration is 3%; Afterwards this mixture is loaded in a nozzle component of many shower nozzles 3D printing device;
C) using vascular endothelial cell and neuron as passage cell; Vascular endothelial cell is mixed with sodium alginate soln, loads in second nozzle component of 3D printing device; Neuron is mixed with cell culture fluid (DMEM), loads in the 3rd nozzle component of 3D printing device;
D) the different nozzle components of many shower nozzles 3D printing device are controlled by computer, the shaping of the hepatoma carcinoma cell/sodium alginate of control subject structure; The channel formation of synchronous or asynchronous controlling vascular endothelial cell/sodium alginate, obtains vascular system passage; Synchronous or asynchronous controlling neuron/DMEM, is located with in access opening, obtains nervous system passage; And adopt CaCl
2sodium alginate soln is cross-linked and obtains Sodium Alginate Hydrogel Films, successively pile up, obtain described containing multichannel twin-channel imitative liver tumor structure;
E) by sample stage as electromagnetism training devices of the imitative liver tumor structure for preparing;
F) positive pole of electromagnetism training devices and the metal plate of negative pole imitated the both sides of cardiac structure but do not contact;
G) be connected with impulsive motion bioreactor mozzle by the vascular system of multichannel sample, start pulsation culture systems, and the flow velocity controlling wherein culture fluid is 10mm/s, action time 20min;
H) just the electric field generation device of electromagnetism training devices is energized, and controls size of current 5mA, direct current;
I) complete electric field to after the training of bionical liver tumor structure, proceeded In vitro culture.
Embodiment 11: utilize many shower nozzles 3D impact system to prepare a kind of containing the three-channel imitative cartilage structure of blood vessel, nerve and immune system, and this imitative cartilage structure is trained in the electric field.
A) threedimensional model containing multichannel imitative cartilage structure described in Computer Design is utilized;
B) using chondrocyte as host cell, mix with the fibrinogen solution configured, fibrinogen solution mass body volume concentrations is 3%; Afterwards this mixture is loaded in a nozzle component of many shower nozzles 3D printing device;
C) using vascular endothelial cell, smooth muscle cell, neuron and T lymphocyte as passage cell; Vascular endothelial cell and smooth muscle cell are mixed with sodium alginate soln, loads in second nozzle component of 3D printing device; Neuron is mixed with sodium alginate soln, loads in the 3rd nozzle component of 3D printing device; T lymphocyte is mixed with sodium alginate soln, loads in the 4th nozzle component of 3D printing device;
D) controlled the different nozzle components of many shower nozzles 3D printing device by computer, the chondrocyte/fibrinogenic shaping of control subject structure, and adopt thrombin to make fibrinogen gel; The channel formation of synchronous or asynchronous controlling capillary endothelial cell and smooth muscle cell/sodium alginate, obtains vascular system passage; The channel formation of synchronous or asynchronous controlling neuron/sodium alginate, obtains nervous system passage; The channel formation of synchronous or asynchronous controlling T lymphocyte/sodium alginate, obtains immune system passage; Adopt CaCl
2make sodium alginate soln gelation obtain Sodium Alginate Hydrogel Films, successively pile up, obtain described containing multichannel three-channel imitative cartilage structure.
E) by sample stage as electromagnetism training devices of the imitative cartilage structure for preparing;
F) positive pole of electromagnetism training devices is connected with imitative cartilage structure with the metal probe of negative pole, makes imitative cartilage structure become a circuit part;
G) just the electric field generation device of electromagnetism training devices is energized, and is 2mA by control size of current, exchanges;
H) complete electric field to after the training of imitative cartilage structure, proceeded In vitro culture.
Claims (13)
1., containing a biomimetic features for passage, it is characterized in that: described biomimetic features comprises main structure body (101) and at least one passage, described passage is distributed in main structure body (101); Described main structure body is the mixture of natural polymer hydrogel and cell composition, and described cell concentration is 10-10
8individual/mL, this cell is at least one in embryonic stem cell, adult stem cell, adult cell, cancerous cell and induced multi-potent stem cells; Described passage is single channel or branched bottom (105), and passage is the one or more combination in two ends through hole (103), two ends blind hole (106) and blind hole one end, one end through-hole structure (104); Position relationship between passage is crossing, parallel, conllinear or antarafacial; Be distributed with cell in described channel outer wall, vias inner walls or access opening, cell is at least two kinds in nervous system seed cell, vascular system seed cell and immune system seed cell.
2. a kind of biomimetic features containing passage as claimed in claim 1, is characterized in that: the cross section of described passage is circular, oval, polygon or irregular geometric figures, and cross sectional area is 100 μm
2-1cm
2.
3. a kind of biomimetic features containing passage as claimed in claim 1, is characterized in that: described nervous system seed cell is at least one in neuron and neurogliocyte; Described blood vessel seed cell is at least one in endotheliocyte, smooth muscle cell and fat stem cell; Described immune system seed cell is at least one in lymphocyte and inherent immunity cell.
4. a kind of biomimetic features containing passage as claimed in claim 1, it is characterized in that: described natural polymer hydrogel is at least one in sodium alginate, collagen, matrigel, dextrose, chitosan, gelatin and Fibrinogen, the mass body volume concentrations of this hydrogel is 0.1 ~ 20%.
5. a kind of biomimetic features containing passage as claimed in claim 1, is characterized in that: be compounded with at least one in cell cryopreservation agent, cell growth factor, medicine, anticoagulant and magnetic nanoparticle in described natural polymer hydrogel; Described cell cryopreservation agent is at least one in dimethyl sulfoxide, glycerol and dextrose; Described cell growth factor is at least one in VEGF, basic fibroblast growth factor, hepatocyte growth factor, human blood platelets derived growth factor and transforming growth factor; Described medicine is at least one in antitumor drug and viral vaccine; Described anticoagulant is at least one in heparin and paclitaxel; Described magnetic nanoparticle is at least one in ferrite particle, metal mold granule and nitrided iron granule.
6., containing an electromagnetic force training devices for the biomimetic features of passage, it is characterized in that: described device comprises electric field generation system (601), magnetic field generation system, sample stage (603) and fixed platform (605); Described magnetic field generation system and sample stage (603) are installed in fixed platform (605); Described electric field system (601) system is containing positive pole and negative pole; Described sample stage (603) is provided with guide rail, and described positive pole and negative pole are arranged on guide rail respectively by slide block; Described sample stage (603) is positioned at magnetic field generation system; Described magnetic field generation system divides and comprises support (602) and rotatable annulus (608), described rotatable annulus (608) is arranged in the hollow structure of support (602), is placed with dismountable S pole (606) and N pole (607) in the upper radial symmetric of rotatable annulus (608).
7. the electromagnetic force training devices of a kind of biomimetic features containing passage as claimed in claim 6, is characterized in that: described device also comprises pulsation culture systems (701); Described pulsation culture systems is installed in fixed platform (605); Described pulsation culture systems (701) comprises pulsation system motor (702), pulsation system guide rail-slide block mechanism (703), culture fluid supply syringe (704), check valve (705), mozzle (706) and culture jar (708); Described pulsation system motor (702) is connected with pulsation system guide rail-slide block mechanism (703) by crank; Described pulsation system guide rail-slide block mechanism (703) and culture fluid supply syringe (704) and are connected.
8. the electromagnetic force training devices of a kind of biomimetic features containing passage as claimed in claims 6 or 7, is characterized in that: described positive pole and negative pole are the one in metal plate, plain conductor or metal probe.
9. the electromagnetic force training devices of a kind of biomimetic features containing passage as claimed in claims 6 or 7, is characterized in that: described S pole and the N one very in permanent magnet or electric magnet.
10. adopt the method that device is as claimed in claim 6 trained the biomimetic features containing passage, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) start the electric field generation system of electromagnetism training devices, remove N level and the S level of magnetic field generation system; The biomimetic features containing passage is made to be arranged in electric field; Electric current, for being greater than 0, is less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly;
C) complete electric field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
11. 1 kinds of methods adopting device as claimed in claim 6 to train the biomimetic features containing passage, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) close the electric field generation system of electromagnetism training devices, start magnetic field generation system, S pole and N pole are rotated with rotatable annulus, controlling magnetic induction for being greater than 0, being less than or equal to 5T;
C) complete magnetic field to after the training of the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
12. 1 kinds of methods adopting device as claimed in claim 6 to train the biomimetic features containing passage, it is characterized in that, described method comprises the steps:
A) biomimetic features containing passage prepared is immersed in the cell culture fluid containing suspension cell, and be placed on the sample stage of electromagnetic force training devices;
B) start electric field generation system and the magnetic field generation system of electromagnetism training devices simultaneously, make the biomimetic features containing passage be arranged in Electric and magnetic fields; Controlling electric current for being greater than 0, being less than or equal to 50mA; Voltage, for being greater than 0, is less than or equal to 50V; The sense of current adopt exchange, direct current or both be used alternatingly; Controlling magnetic induction for being greater than 0, being less than or equal to 5T;
C), after completing the training of Electric and magnetic fields simultaneously to the biomimetic features containing passage, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
13. a kind of methods to training containing the biomimetic features of passage according to claim 10,11 or 12, it is characterized in that: start pulsation culture systems (701), described pulsation culture systems (701) comprises pulsation system motor (702), pulsation system guide rail-slide block mechanism (703), culture fluid supply syringe (704), check valve (705), mozzle (706) and culture jar (708); Described pulsation system motor (702) is connected with pulsation system guide rail-slide block mechanism (703) by crank; Described guide rail-slide block mechanism and culture fluid supply syringe (704) and are connected; By mozzle (706), described biomimetic features is connected with pulsation culture systems; Culture fluid is made to carry out one-way flow between mozzle (706) and the biomimetic features containing passage; Flow rate of liquid, for being greater than 0, is less than 30cm/s, the effect of-electric field that carries out pulsing, pulsation-magnetic field or pulsation-electromagnetic complex field; After completing the training to described biomimetic features, by this biomimetic features cryopreservation, or proceed In vitro culture or be directly used in organ transplantation.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410535181.8A CN104306083B (en) | 2014-10-11 | A kind of biomimetic features containing passage and electromagnetic force training devices thereof and method | |
| PCT/CN2014/089771 WO2016054847A1 (en) | 2014-10-11 | 2014-10-29 | Bionic structure containing channels and electromagnetic force training device and method therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410535181.8A CN104306083B (en) | 2014-10-11 | A kind of biomimetic features containing passage and electromagnetic force training devices thereof and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104306083A true CN104306083A (en) | 2015-01-28 |
| CN104306083B CN104306083B (en) | 2017-01-04 |
Family
ID=
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104658395A (en) * | 2015-02-15 | 2015-05-27 | 清华大学 | Heart simulation structure as well as forming method and special mold thereof |
| CN107937339A (en) * | 2016-10-13 | 2018-04-20 | 中国科学院大连化学物理研究所 | A kind of external model method for building up of gestational period alcohol exposure to fetus brain damage |
| CN107937344A (en) * | 2016-10-13 | 2018-04-20 | 中国科学院大连化学物理研究所 | A kind of method for the brain growth that hiPSCs sources are realized using macaroni yarn as carrier |
| CN108369224A (en) * | 2015-10-02 | 2018-08-03 | 小利兰·斯坦福大学托管委员会 | It is sorted using the biology and abiological constitution of magnetic suspension |
| CN111246941A (en) * | 2017-10-02 | 2020-06-05 | 哈利法科学技术大学 | Microfluidic device for generating lymph nodes in vitro |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050222591A1 (en) * | 2004-03-30 | 2005-10-06 | Peter Gingras | Medical device |
| US20050281856A1 (en) * | 2004-05-10 | 2005-12-22 | Mcglohorn Jonathan | Implantable biostructure comprising an osteoconductive member and an osteoinductive material |
| CN101084026A (en) * | 2004-06-30 | 2007-12-05 | 伊利诺伊州大学理事会 | Poly (ethylene glycol) - diacrylate (pegda) - crosslinked comprising adipogenic mesenchymal stem cells |
| CN101148656A (en) * | 2006-09-18 | 2008-03-26 | 中国人民解放军军事医学科学院野战输血研究所 | Construction method for tissue engineering liver unit and tissue engineering liver unit |
| CN101920046A (en) * | 2005-02-23 | 2010-12-22 | Hi-Lex株式会社 | Medical material, artificial tooth root and method of producing material for clinical use |
| CN101954118A (en) * | 2010-10-13 | 2011-01-26 | 同济大学 | Method for preparing aqueous gel/nano hydroxyapatite composite scaffold for hard tissue repair |
| CN103767804A (en) * | 2014-01-20 | 2014-05-07 | 清华大学 | Vascularizing tissue structure with microfluid passage and preparation method thereof |
| CN103908698A (en) * | 2014-04-15 | 2014-07-09 | 中国人民解放军第四军医大学 | Superparamagnetic iron oxide nano-particle-collagen-chitosan magnetic degradable nerve conduit and preparation method thereof |
| CN204181751U (en) * | 2014-10-11 | 2015-03-04 | 清华大学 | A kind of device biomimetic features containing passage being carried out to electromagnetic force training |
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050222591A1 (en) * | 2004-03-30 | 2005-10-06 | Peter Gingras | Medical device |
| US20050281856A1 (en) * | 2004-05-10 | 2005-12-22 | Mcglohorn Jonathan | Implantable biostructure comprising an osteoconductive member and an osteoinductive material |
| CN101084026A (en) * | 2004-06-30 | 2007-12-05 | 伊利诺伊州大学理事会 | Poly (ethylene glycol) - diacrylate (pegda) - crosslinked comprising adipogenic mesenchymal stem cells |
| CN101920046A (en) * | 2005-02-23 | 2010-12-22 | Hi-Lex株式会社 | Medical material, artificial tooth root and method of producing material for clinical use |
| CN101148656A (en) * | 2006-09-18 | 2008-03-26 | 中国人民解放军军事医学科学院野战输血研究所 | Construction method for tissue engineering liver unit and tissue engineering liver unit |
| CN101954118A (en) * | 2010-10-13 | 2011-01-26 | 同济大学 | Method for preparing aqueous gel/nano hydroxyapatite composite scaffold for hard tissue repair |
| CN103767804A (en) * | 2014-01-20 | 2014-05-07 | 清华大学 | Vascularizing tissue structure with microfluid passage and preparation method thereof |
| CN103908698A (en) * | 2014-04-15 | 2014-07-09 | 中国人民解放军第四军医大学 | Superparamagnetic iron oxide nano-particle-collagen-chitosan magnetic degradable nerve conduit and preparation method thereof |
| CN204181751U (en) * | 2014-10-11 | 2015-03-04 | 清华大学 | A kind of device biomimetic features containing passage being carried out to electromagnetic force training |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104658395A (en) * | 2015-02-15 | 2015-05-27 | 清华大学 | Heart simulation structure as well as forming method and special mold thereof |
| CN104658395B (en) * | 2015-02-15 | 2017-04-19 | 清华大学 | Heart simulation structure as well as forming method and special mold thereof |
| CN108369224A (en) * | 2015-10-02 | 2018-08-03 | 小利兰·斯坦福大学托管委员会 | It is sorted using the biology and abiological constitution of magnetic suspension |
| CN107937339A (en) * | 2016-10-13 | 2018-04-20 | 中国科学院大连化学物理研究所 | A kind of external model method for building up of gestational period alcohol exposure to fetus brain damage |
| CN107937344A (en) * | 2016-10-13 | 2018-04-20 | 中国科学院大连化学物理研究所 | A kind of method for the brain growth that hiPSCs sources are realized using macaroni yarn as carrier |
| CN107937339B (en) * | 2016-10-13 | 2021-06-11 | 中国科学院大连化学物理研究所 | In-vitro model establishment method for fetal brain injury caused by alcohol exposure in gestation period |
| CN107937344B (en) * | 2016-10-13 | 2021-06-11 | 中国科学院大连化学物理研究所 | Method for realizing brain development of hiPSCs source by taking hollow fibers as carrier |
| CN111246941A (en) * | 2017-10-02 | 2020-06-05 | 哈利法科学技术大学 | Microfluidic device for generating lymph nodes in vitro |
| US11478798B2 (en) | 2017-10-02 | 2022-10-25 | Khalifa University of Science and Technology | Microfluidic device for generating an in vitro lymph node |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016054847A1 (en) | 2016-04-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhao et al. | Review on the vascularization of organoids and organoids-on-a-C hip | |
| Leberfinger et al. | Bioprinting functional tissues | |
| Zhang et al. | 3D bioprinting: a novel avenue for manufacturing tissues and organs | |
| Jang et al. | Biomaterials-based 3D cell printing for next-generation therapeutics and diagnostics | |
| CN111065422A (en) | Method of making a multilayered tubular tissue construct | |
| CN106434562B (en) | Brain tumor in-vitro model for three-dimensional biological printing and construction method thereof | |
| ES2589309T3 (en) | Manufacture of vascularized tissue using microfabricated two-dimensional molds | |
| CN105985925B (en) | A kind of global function artificial organs fitting body and its preparation and cultural method | |
| CN101492655B (en) | Vascularized fat depot based on partition and construction method thereof | |
| CN110403731B (en) | Tissue engineering bionic liver lobe structure based on living cell 3D printing and preparation method | |
| CN106421916A (en) | Tissue engineering skin and preparation method thereof | |
| Zhang et al. | Organ-level vascularization: The Mars mission of bioengineering | |
| WO2003078586A2 (en) | Vascularized tissue for transplantation | |
| Tabatabaei Rezaei et al. | Recent advances in organ‐on‐chips integrated with bioprinting technologies for drug screening | |
| Liu et al. | Vascularization of engineered organoids | |
| CN110607271B (en) | Preparation method of in vitro vascularized 3D tissue based on micromachining technology | |
| Jeyaraj et al. | Vascularisation in regenerative therapeutics and surgery | |
| Cho et al. | 3D Bioprinting | |
| Li et al. | Advances of 3D printing in vascularized organ construction | |
| Visconti et al. | Cardiovascular tissue engineering I. Perfusion bioreactors: a review | |
| US20050003524A1 (en) | Module for the culture and/or preservation of microorganisms and the utilization of its metabolic performance | |
| CN112166179A (en) | Systems and methods for multi-channel vasculature | |
| CN116004388A (en) | Microfluidic chip and in-vitro three-dimensional organoid model construction method | |
| CN104306083A (en) | Bionic structure with passageways as well as electromagnetic force training device and electromagnetic force training method thereof | |
| CN104306083B (en) | A kind of biomimetic features containing passage and electromagnetic force training devices thereof and method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170104 Termination date: 20171011 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |