CN104207859B - Rotation method of piling is utilized to prepare method and the special equipment of histoorgan - Google Patents
Rotation method of piling is utilized to prepare method and the special equipment of histoorgan Download PDFInfo
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- CN104207859B CN104207859B CN201410473055.4A CN201410473055A CN104207859B CN 104207859 B CN104207859 B CN 104207859B CN 201410473055 A CN201410473055 A CN 201410473055A CN 104207859 B CN104207859 B CN 104207859B
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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
Landscapes
- Health & Medical Sciences (AREA)
- Cardiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Materials For Medical Uses (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Utilize rotation method of piling to prepare method and the special equipment of histoorgan, belong to tissue engineering technique field.This invention is based on soluble core technique and cell assembling technology, by various cell growth factor, inhibitive factor or medicine are compound in the hydrogel material of cross-linking/polymerization, from shower nozzle extrusion thread hydrogel crosslinked/polymerization, synthesis macromolecule is combined, it is wound around after culture fluid carrying out washing treatment and is deposited on rotary inner core, hydrogel containing dissimilar seed cell and adhesion endotheliocyte, the hydrogel of neurocyte forms main part and blood vessel respectively, nervous system, after shaping molten except inner core and obtain that there is corkscrew vessels and neural histoorgan after cultivating a period of time.Present invention process is simple and is readily formed blood vessel and nervous system, being particularly well-suited to shape the histoorgan with inner chamber that tradition organizational project is difficult to shape, the histoorgan of preparation can be pasted at disease damage histoorgan or replace disease damage histoorgan in directly transplanting body.
Description
Technical field
The present invention relates to the use of rotation method of piling and prepare method and the special equipment of histoorgan, belong to organizational project neck
Territory.
Background technology
Human body disease damage tissue and the reparation of organ and replacement are the science frontiers attracted people's attention this century, are also one and need badly
The sciences problems solved.Owing to biological tissue organ transplantation exists donor shortage and the intrinsic drawback such as immunologic rejection risk is big,
It is difficult to become the effective way solving this problem.Accordingly, the organizational project for the purpose of improving this type of illness treatment level
(Tissue Engineering) arises at the historic moment, and organizational project is intended to the three-dimensional structure of external structure cell and material, passes through
Develop into, after suitable cultivation and training, the transplantation substitute having with natural tissues similar structures and function.In recent years, group
The focus of weaver's journey research the most both at home and abroad, quickly grows and achieves many achievements, but for its product applications,
Currently mainly concentrate on a few fields such as the relatively simple bone tissue engineer of structure and skin tissue engineering.Trace it to its cause,
The very important point is exactly, and the vitro construction method (such as Electrospun, die casting etc.) of existing organizational project structure is difficult to structure
Building and have the complicated blood vessel network structure that natural tissues is similar, the cell survival rate causing construction inner is low, therefore, it is difficult to
Realize the structure of bulk tissue.
Traditional organizational project follows the Constructed wetlands of " from top to bottom " of " cell-scaffold is combined ", is i.e. shaped support,
Again on cell seeding to support.But, when stent size and complexity bring up to a certain degree, stent forming and cell are multiple
The difficulty closed will be greatly increased.Cell three-dimensional controlled group packing technique (Wang X H, Yan Y N, Zhang R J.Rapid
prototyping as a tool for manufacturing bio-artificial livers.Trends in
Biotechnology, 2007,25 (11): 505-513.), i.e. cell 3D printing technique, it then follows " support-cell " integration structure
The thinking built.During cell printing, cell (or cell aggregation) and colloidal sol (presoma of hydrogel) be concurrently placed at or
The celliferous culture fluid of person is individually placed in the shower nozzle of printer, computer control the deposition position containing cell drop, is referring to
Fixed position pointwise prints, and continues to print another layer on the basis of having printed one layer, and the three-dimensional many cells of the formation that is layering coagulate
Colloid system.The feature of cell printing technology is the uniformity that can ensure that cell density and distribution, and can realize multiple carefully
The common of born of the same parents assembles and fixed point arrangement.But oxygen and effective transportation problem of nutrient substance, i.e. " vascularization problem " remain
The method builds the bottleneck problem of transplantation complicated tissue organ.
The existing histoorgan by cell 3D printing-forming is all to pile up according to plane in layer, because of
Its inherent characteristics shaped, the shaping of microchannel (blood vessel) depends on the character of forming accuracy and material.And in for having
The shaping of cavity configuration histoorgan, the cell printing technology relying on plane to pile up inevitably needs to increase in its forming process
Add support or rotate forming table or shower nozzle orientation, considerably increasing design and the difficulty shaped, the most helpless.
Chinese patent literature CN1654028 proposes a kind of method utilizing molten core legal system tubing network, first with water
Dissolubility, the material of inanimate object toxicity manufacture the inner core of support, are then coated in by biocompatible materials on support inner core, through wind
Use distillation water-soluble except support inner core after Gan.The method can only the better simply grid shape stent of shaped structure, for having complicated blood
The shaping of the histoorgan of pipe is helpless.
Summary of the invention
It is an object of the invention to overcome traditional organizational project is difficult to build and there is complicated blood vessel and neuromechanism
Cavity type histoorgan, it is provided that a kind of method and special equipment preparing histoorgan based on rotation method of piling.The method is carefully
On the basis of born of the same parents' controlled group packing technique and soluble core technique, it is therefore intended that the preparation method of a kind of new histoorgan is provided, it is simple to
Preparation has spiral net-shaped blood vessel and neural histoorgan, is particularly useful for making the histoorgan with inner-cavity structure.
Technical scheme is as follows:
Rotation method of piling is utilized to prepare the process of histoorgan, it is characterised in that the special equipment of the present invention includes propping up
Support arrangement, many nozzle components, motion, forming device and control system；Described support means includes supporting leg and base plate；
Described motion uses guide rail slide block mechanism, including X direction guiding rail, X to slide block, Y-direction guide rail, the first Y-direction slide block, the second Y-direction
Slide block, Z-direction guide rail, Z-direction slide block, described X direction guiding rail is located on base plate, and Z-direction guide rail is fixed in X on slide block, and Y-direction guide rail is affixed
On Z-direction slide block；Described many nozzle components at least contain six shower nozzles, including at least two squash type shower nozzle and four spray-types
Shower nozzle, the nozzle inside diameter of two squash type shower nozzles is respectively 200 μm～1mm and 50 μm～500 μm, the net of spray-type spray nozzle
Bore dia is 50 μm～100 μm, and shower nozzle divides two groups to be separately mounted on the first Y-direction slide block and the second Y-direction slide block；Described shaping fills
Putting and include forming containers, support and motor, forming containers is arranged on base plate and is in the bottom of nozzle component, and support is arranged on
In forming containers and it is connected with motor output shaft.
Special equipment of the present invention is characterized in that: the structure of described support is that two ends are cylindrical, middle for prism
Shape, material is rustless steel or titanium alloy.
Special equipment of the present invention is characterized in that: have on described first Y-direction slide block and the second Y-direction slide block with
It is respectively mounted the first little cunning on first little guide rail of Y-direction guide rail parallel and the second little guide rail, the first little guide rail and the second little guide rail
Block) and the second small slide block, the first small slide block and the second small slide block on have can the hole of stationary nozzle.
Utilize and rotate the method that method of piling prepares histoorgan, it is characterised in that the method comprises the following steps:
1) prepared by material:
Preparation 0.2～20% (w/v) hydrogel solution, 1～10% (w/v) cross-linking agent/polymer fluid, 0.1～10% (w/v)
Synthesis macromolecular solution, 0.2～10% (w/v) gelatin solution and cell culture fluid；Separate or induction seed cell, plant careful
Born of the same parents' density is 1 × 103～1 × 106Individual/mL, is mixed seed cell and hydrogel solution by 1～9:9～1 volume ratio and contains
The hydrogel solution of seed cell；Separating or inducing nerve cell and endotheliocyte, and make cell suspension, neurocyte hangs
Liquid and endotheliocyte suspension density are 1 × 103～1 × 108Individual/mL；
2) prepared by inner core
When preparation has the histoorgan of inner chamber, according to the cavity shape of histoorgan to be prepared, use three-dimensional drawing
Its threedimensional model of software design, makes the mould with this inner-cavity structure, when the histoorgan of forming solid by quick shaping method
Time, mould is the cylinder of diameter thick 8～10mm；Support is put into mould central authorities, after adding gelatin solution in mould, reduces temperature
Degree makes gelatin solidify to 8～10 DEG C, opens mould and obtains the histoorgan inner core of center band support, at the histoorgan of belt supporting frame
Coat hydrogel outside inner core, and carry out cross-linking/being polymerized with cross-linking agent/polymer fluid；
3) shape
Hydrogel solution containing seed cell and the most celliferous hydrogel solution are respectively charged into the first squash type shower nozzle
With in the second squash type shower nozzle, cross-linking agent/polymer fluid, neurocyte suspension, endotheliocyte suspension and synthesis macromolecular solution
It is respectively charged in the 4th spray-type shower nozzle, the second spray-type shower nozzle, the first spray-type shower nozzle and the 3rd spray-type shower nozzle, is shaping
Container content enters cell culture fluid, is connected in support is arranged on forming containers and with motor output shaft, by organizer to be formed
After the model of official imports control system, start former, start stack shaping；In forming process, support does around Y negative sense and revolves
Transhipment is dynamic, and shower nozzle does translational motion, and when shaping the main part of histoorgan, the first squash type shower nozzle moves to specify position
The extrusion hydrogel containing seed cell is wrapped on inner core, simultaneously equipped with the 4th spray-type shower nozzle motion of cross-linking agent/polymer fluid
Dead astern to the first squash type shower nozzle sprays cross-linking agent/polymer fluid and cross-links it/be polymerized；When shaping blood vessel, the
Two squash type shower nozzles move to specify position to extrude the most celliferous hydrogel and are wrapped on inner core, the 4th spray-type shower nozzle simultaneously
Moving to the second squash type shower nozzle dead astern, the first spray-type shower nozzle and the 3rd spray-type shower nozzle move to the second squash type respectively
Shower nozzle dead ahead, under the inner core rotated drives, the new hydrogel being wound around moves to hand under the 4th spray-type shower nozzle successively
Connection/polymerization, washs through culture fluid, carries out spraying endotheliocyte to the first spray-type shower nozzle, the 3rd spray-type shower nozzle
With composite synthesis macromolecular solution；As body and the spirit through time, the second squash type shower nozzle move to specify position extrusion the most celliferous
Hydrogel is wrapped on inner core, and the 4th spray-type shower nozzle moves to the second squash type shower nozzle dead astern, the second spray-type spray simultaneously
Head and the 3rd spray-type shower nozzle move to the second squash type shower nozzle dead ahead respectively, under the inner core rotated drives, are newly wound around
Hydrogel moves to carry out cross-linking/being polymerized under the 4th spray-type shower nozzle successively, washs through culture fluid, to the second spray-type
Carry out under shower nozzle, the 3rd spray-type shower nozzle spraying neurocyte and composite synthesis macromolecule；So the most repeatedly it is wound around and piles up until shape
Become the histoorgan precursor containing inner core；
4) subsequent treatment
Histoorgan precursor containing inner core is placed in calorstat cultivation, molten except taking out support, shape after cultivating after inner core
Becoming rotational-like histoorgan, nerve and blood vessel are the most therethrough.
The method preparing histoorgan of the present invention, it is characterised in that: described seed cell is for having differentiation capability
Stem cell or there is the adult cell of physiologically active；Described stem cell is that fat stem cell, embryonic stem cell, blood are dry thin
Born of the same parents, bone marrow stem cell, induction type versatile stem cell, described adult cell is adipose cell, hepatocyte, bladder cells, lymph
Cell, myocardial cell or nephrocyte；Described hydrogel for be compounded with gelatin, collagen, matrigel, carrageenan, chitosan, agar,
One or more sodium alginate or fibrinogen solution in hyaluronic acid, matrigel, elastin laminin and laminin；Described
Hydrogel can be combined various cell cryopreservation agent, cell growth factor, medicine and anticoagulant；Cell cryopreservation agent is that dimethyl is sub-
One or more complex in sulfone (DMSO), glycerol and dextrose；Described cell growth factor is vascular endothelial growth factor
Son (VEGF), basic fibroblast growth factor (b-FGF), hepatocyte growth factor (HGF), human blood platelets derived growth because of
One or more complex in son (PDGF-BB) and transforminggrowthfactor-β1 (TGF-β 1)；Described medicine is antineoplastic agent
Thing, this antitumor drug is astragalus polysaccharides, cis diamino dichloro network platinum (DDP), mitomycin (MMC), 5-fluorouracil (5-
FU), one or more complex in Claritin, antibiotic, viral vaccine；Described anticoagulin is heparin, purple
One or more complex in China fir alcohol；The solute of described synthesis macromolecular solution be polyurethane, polycaprolactone, Merlon,
One or more in Polyethylene Glycol, Poly(D,L-lactide-co-glycolide, polyester and polyhydroxy acid ester, solvent is tetrem two
Alcohol or 1,4-dioxane.
The present invention compared with prior art, has the advantage that and the technique effect of salience:
1. molten core method and 3D Method of printing are combined by the present invention, can shape the tool that tradition 3D Method of printing is difficult to shape
There is the histoorgan of inner chamber；
The use of the most shower nozzles can realize various kinds of cell heterogeneous body and shape and surface compound treatment, meets natural tissues device
Official's requirement to various kinds of cell；
3. blood vessel (neural) generation type at hydrogel surface sprinkling endothelium (neural) cell can be formed with complicated knot
The blood vessel (neural) of structure, the thin film that synthesis macromolecular solution is formed improves endothelium (neural) cell number that hydrogel surface adheres to
Mesh, effectively facilitates the formation of blood vessel (neural)；
4. in forming process, cell culture fluid can be that cells with nutrient material again can be with the crosslinking on detergent gel surface
Organic solvent in agent/polymer fluid and synthesis macromolecule, reduces its damage to cell.
Accompanying drawing explanation
Fig. 1 is the special equipment three dimensional structure sketch of the present invention.
Fig. 2 is the three dimensional structure sketch of the first Y-direction slide block.
Fig. 3 is the histoorgan forming process schematic diagram of the present invention.
Fig. 4 is the histoorgan forming technology flow chart of the present invention.
Fig. 5 a is the most celliferous hydrogel；Fig. 5 b is the hydrogel after spraying endothelium (neural) cell；Fig. 5 c is for spraying
Hydrogel after synthesis macromolecular solution.
Fig. 6 is the structural representation of the histoorgan of the present invention.
In figure: 101 base plates；102 forming containers；103 supports；104 first spray-type shower nozzles；105 second spray-types
Shower nozzle；106 Z-direction slide blocks；107 Y-direction guide rails；108 the 3rd spray-type shower nozzles；109 first small slide blocks；110 first little lead
Rail；111 first Y-direction slide blocks；112 second Y-direction slide blocks；113 first squash type shower nozzles；114 second squash type shower nozzles；115‐
4th spray-type shower nozzle；116 second small slide blocks；117 second little guide rails；118 Z-direction guide rails；119 electric rotating machines；120 X to
Slide block；121 X direction guiding rails；401 inner cores；402 nozzle components；403 materials；404 water-setting collodion silks；405 culture fluid；601‐
Main part；602 blood vessels (neural)；603 inner chambers.
Detailed description of the invention
The present invention is further described with embodiment below in conjunction with the accompanying drawings.
Fig. 1 is that the rotation method of piling that utilizes that the present invention provides prepares the special equipment of histoorgan, and this equipment includes spraying more
Head assembly, motion, forming device and control system (picture).Described motion is realized by slide block guide rail (groove),
Including X direction guiding rail 121 and X to slide block 120, Y-direction guide rail 107 and first Y-direction slide block the 111, second Y-direction slide block 112, Z-direction guide rail
118 and Z-direction slide block 106 and the first small slide block the 109, second small slide block 116 and first little guide rail the 110, second little guide rail 117；
Described X direction guiding rail 121 is located on base plate 101；Described Z-direction guide rail 118 is fixed in X on slide block 120；Described Y-direction guide rail 107 is solid
It is connected on Z-direction slide block 106；Described first little guide rail 110 and the second little guide rail 117 are separately mounted to the first Y-direction slide block 111 and
On two Y-direction slide blocks 112；Described many nozzle components at least contain six shower nozzles, including at least two squash type shower nozzle and four sprays
Spill formula shower nozzle, divide two groups and be separately fixed on two parallel Y-direction slide blocks, first spray-type shower nozzle the 104, second spray-type shower nozzle
105 and the 3rd spray-type shower nozzle 108 be arranged on the first Y-direction slide block 111, wherein the first spray-type shower nozzle 104 and second sprays
The shower nozzle that formula shower nozzle 105 is arranged on the first Y-direction slide block 111 is fixed in groove, and the 3rd spray-type shower nozzle 108 is arranged on the first little cunning
The shower nozzle of block 109 is fixed on groove；4th spray-type shower nozzle 115 and at least two squash type shower nozzle, the first squash type shower nozzle 113 He
Second squash type shower nozzle 114, is arranged on the second Y-direction slide block 112, wherein the first squash type shower nozzle 113 and the spray of the second squash type
114 shower nozzles being fixed on the second Y-direction slide block 112 are fixed on groove, and the 4th spray-type shower nozzle 115 is arranged on the second small slide block 116
Shower nozzle fix on groove, the nozzle inside diameter of the first squash type shower nozzle 113 and the second squash type shower nozzle 114 is respectively 200 μm 1mm
With 50 μm 500 μm, the mesh diameter of four spray-type spray nozzles is 50 μm 100 μm；Described forming device includes into and describing
Device 102, support 103 and motor 119, forming containers 102 is arranged on the underface of base plate 101 upper nozzle assembly, dismountable
In support 103 is arranged on forming containers 102 and being connected with the output shaft of motor 119, described support 103 is strong for having certain mechanics
The solid material to cytotoxic of degree, such as rustless steel or titanium alloy；Described control system is responsible for controlling nozzle component
D translation motion and the rotary motion of support.
Fig. 2 is the three dimensional structure sketch of the first Y-direction slide block, described first Y-direction slide block has two and is used for fixing shower nozzle
Hole and one can allow spray cephalomotor macropore, macropore both sides are provided with the first little guide rail 110, and the first small slide block 109 can be led along little
Rail moves, and driving the 3rd spray-type shower nozzle 108 thereon is the first spray-type shower nozzle 104 second spray-type shower nozzle 105 work in turn
Making, the second Y-direction slide block structure and the first Y-direction slide block are similar to.
Fig. 3 is the flow chart rotating method of piling formation tissue organ that utilizes of the present invention, mainly includes the preparation before shaping
The three big key steps such as work, histoorgan forming precursor and subsequent operation.
Preparation before shaping includes the preparation of material and the making of inner core, preparation 0.2～20% (w/v) water-setting peptization
Liquid, 1～10% (w/v) cross-linking agent/polymer fluid, 0.1～10% (w/v) synthesis macromolecular solution, 0.2～10% (w/v) bright
Sol solution and cell culture fluid；According to the type of histoorgan to be formed, separate from a patient and there is differentiation capability
Autologous stem cells, such as fat stem cell, embryonic stem cell, blood stem cell, bone marrow stem cell, induction type versatile stem cell,
Or there is physiologically active adult cell, as adipose cell, hepatocyte, bladder cells, lymphocyte, myocardial cell or nephrocyte are made
For seed cell, and making cell suspension, seed cell density is 1 × 103～1 × 108Individual/mL, by seed cell suspension with go out
Hydrogel solution after bacterium is 1～9:9～1 uniformly to mix by volume, is prepared as the aqueous gel mixture containing seed cell；Institute
State hydrogel for can composite gelatin, collagen, matrigel, carrageenan, chitosan, agar, hyaluronic acid, matrigel, elastin laminin,
In laminin, one or more sodium alginate or fibrinogen solution, can be combined various cell cryopreservation in described hydrogel
Agent, cell growth factor, medicine, anticoagulant.Cell cryopreservation agent is in dimethyl sulfoxide (DMSO), glycerol, dextrose
Plant or multiple complex.Described cell growth factor be endothelial cell growth factor (ECGF) (as VEGF (VEGF),
Basic fibroblast growth factor (b-FGF), hepatocyte growth factor (HGF), human blood platelets derived growth factor (PDGF-
BB), one or more complex in transforminggrowthfactor-β1 (TGF-β 1).Described medicine is that antitumor drug is (such as the Radix Astragali
Polysaccharide, cis diamino dichloro network platinum (DDP), mitomycin (MMC), 5-fluorouracil (5-FU), Claritin, antibiotic
One or more complex in (such as penicillin, gibberellins, tetracycline), viral vaccine.Described anticoagulin be heparin,
One or more complex in paclitaxel；The solute of described synthesis macromolecular solution is specially polyurethane (PU), polycaprolactone
(PCL), the one in Merlon, Polyethylene Glycol, PLGA (PLGA), polyester and polyhydroxy acid ester
Or multiple, solvent is TEG or Isosorbide-5-Nitrae dioxane；Take out the support 103 on forming containers 102 and carry out at sterilizing
Reason, when being formed with the histoorgan of inner chamber, according to the construction features of histoorgan inner chamber to be formed, uses three-dimensional drawing
Its threedimensional model of software design, makes the mould with this inner-cavity structure, stands in mould by support 103 by quick shaping method,
In mould, add gelatin solution, reduce temperature and make gelatin solidify to 10～12 DEG C, obtain in the histoorgan of center band support
Core；When shaping solid tissue organ, inner core is the cylindrical sleeves of thick 5～6mm；Then outside the histoorgan inner core of belt supporting frame
Coat one layer of hydrogel, and carry out cross-linking/being polymerized with cross-linking agent/polymer fluid.
Fig. 4 is the rotation stack shaping process schematic of the present invention, by the hydrogel containing seed cell and the most celliferous
Hydrogel is respectively charged in equipment in the first squash type shower nozzle 113 and the second squash type shower nozzle 114, cross-linking agent/polymer fluid, nerve
Cell suspension and endotheliocyte suspension are respectively charged into the 4th spray-type shower nozzle the 115, second spray-type shower nozzle 105 and the first spray-type
In shower nozzle 104, synthesis macromolecular solution loads in the 3rd spray-type shower nozzle shower nozzle 108, and support 103 is arranged on forming containers
Cell culture fluid 405 is loaded in 102 and in forming containers 102；After the model of histoorgan to be formed is imported control system,
Starting former, start stack shaping, in forming process, support 103 rotates around Y negative sense, and shower nozzle does translation fortune
Dynamic, when shaping histoorgan main part, the first squash type shower nozzle 113 moves to the water specifying position extrusion containing seed cell
Gel is wrapped on inner core, and the 4th spray-type shower nozzle 115 equipped with cross-linking agent/polymer fluid moves to the first squash type shower nozzle simultaneously
The dead astern of 113 sprays cross-linking agent/polymer fluid and cross-links it/be polymerized；When shaping blood vessel, the second squash type shower nozzle
114 move to specify position to extrude the most celliferous hydrogel is wrapped on inner core, and the 4th spray-type shower nozzle 115 moves to simultaneously
Second squash type shower nozzle 114 dead astern, the first spray-type shower nozzle 104 and the 3rd spray-type shower nozzle 108 move to second respectively and squeeze
Pressure type shower nozzle 114 dead ahead, under the inner core rotated drives, the new hydrogel being wound around moves to the 4th spray-type shower nozzle successively
Carry out for 115 times cross-linking/being polymerized, wash through culture fluid, through the first spray-type shower nozzle the 104, the 3rd spray-type shower nozzle 108
Under carry out spraying endotheliocyte and composite synthesis macromolecule；During as shape nerve fibre bundle, the second squash type shower nozzle 114 moves to
Specifying position to extrude the most celliferous hydrogel to be wrapped on inner core, the 4th spray-type shower nozzle 115 moves to the first extruding simultaneously
Formula shower nozzle 113 dead astern, the second spray-type shower nozzle 105 and the 3rd spray-type shower nozzle 108 move to the second squash type shower nozzle respectively
114 dead aheads, under the inner core rotated drives, the new hydrogel being wound around moves to the 4th spray-type shower nozzle successively to be carried out for 115 times
Crosslinking/polymerization, washs through culture fluid, sprays for 108 times through the second spray-type shower nozzle the 105, the 3rd spray-type shower nozzle
Spill neurocyte and composite synthesis macromolecule, be the most repeatedly wound around accumulation, until the histoorgan needed for Cheng Xinging.
Fig. 5 a is the most celliferous hydrogel；Fig. 5 b is the hydrogel after spraying endothelium (neural) cell；Fig. 5 c is for spraying
Hydrogel after synthesis macromolecular solution；When shaping blood vessel (neural), the most celliferous water-setting collodion silk warp of ejection from shower nozzle
After cross-linking/be polymerized and washing, it is sprayed one layer of endothelium (neural) cell and synthesis macromolecular solution successively, forms gel cell
Membrane structure, in follow-up incubation, hydrogel gradually hydrolyzes, and endotheliocyte or nerve growth also connect each other,
Ultimately form blood vessel and neuromechanism.
The structural representation of the histoorgan that Fig. 6 provides for the present invention, described histoorgan is revolving body or nearly revolving body,
It is made up of inner chamber 603 and peripheral entity；Described inner chamber 603 is closed surface that is cylindric, spherical or that surrounded by other curved surfaces
Shape；Described peripheral entity by containing or the most celliferous water-setting collodion silk be wound around accumulation and form；Hydrogel containing seed cell
Silk 602 is wound agent structure, the most celliferous water-setting collodion silk 603 surface-coated endotheliocyte and synthesis macromolecular solution shape
Vascular system, the most celliferous hydrogel surface coating neurocyte and synthesis macromolecular solution is become to form nervous system.
Enumerate several specific embodiment below, to be further appreciated by the present invention:
The preparation in embodiment 1 atrium
(1) prepared by material
Extract fat stem cell (ADSCs), endotheliocyte (ECs), neurocyte, sternzellen and myocardial cell
(CMCs), subculture, obtain cell material, endotheliocyte and neurocyte concentration and be 8 × 107Individual/mL, sternzellen and
Myocardial cell density is 2 × 106Individual/mL；Take 2g sodium alginate and 7g gelatin be placed in 100mL culture fluid and make it be completely dissolved,
Adding the cells frozen storing liquid DMSO of mass body volume concentrations 3% in this mixed liquor, sterilization treatment obtains gelatin sodium alginate water-setting
Glue material；Taking 4g gelatin to be placed in 100mL culture fluid and make it be completely dissolved, sterilization treatment obtains gelatin solution；Preparation 0.05g/
The calcium chloride water of ml is as its cross-linking agent/polymer fluid；Degradable polycarbonate is dissolved in TEG and obtains mass body
Volume concentrations is the synthesis macromolecular solution of 2%, and sterilization treatment is stand-by；Prepare cell culture fluid DMEM and 1% collagen solution.By star
The mixed liquor of shape cell and myocardial cell is mixed to get hydrogel containing cardiac muscle sternzellen with volume ratio for 1:1 with hydrogel
Solution.
(2) prepared by inner core
Shape and structure feature according to atrium inner chamber, with three-dimensional drawing software design three-dimensional fusiform model, with fast rapid-result
Shape method makes the mould with this inner-cavity structure, erects in mould by propping up after sterilizing, adds gelatin solution, fall in mould
Low temperature makes gelatin solidify to 10～12 DEG C, forms the atrium inner core of center band support, and is coated with outside the atrium inner core of belt supporting frame
Cover one layer of collagen solution, and make collagen solidify to form the protecting film of one layer of water insoluble solution；
(3) shape
Spray being respectively charged into the first squash type containing cardiac muscle sternzellen and the hydrogel of DMSO and the most celliferous hydrogel
113 and the first squash type shower nozzle 114, endotheliocyte suspension and neurocyte suspension be respectively charged into the first spray-type shower nozzle 104,
Second spray-type shower nozzle 105, calcium chloride water loads the 4th spray-type shower nozzle 115, and synthesis macromolecular solution loads the 3rd spray
Spill formula shower nozzle 108, in the support of band gelatin inner core is arranged on forming containers 102 and be connected with motor output shaft, describe at one-tenth
Loading the cell culture fluid of 100mL in device 102, whole device is placed in the environment that temperature is 8～10 DEG C, is imported by atrium model
Start forming device after control system, start to shape.
(4) molten core
After shaping, the atrium precursor containing gelatin inner core is placed in the medium-term and long-term preservation of liquid nitrogen or calorstat cultivation, melts
Except inner core, take out support, i.e. obtain the atrium precursor of band inner chamber.
(5) later stage cultivates
Atrium precursor is cultivated, in incubation, in the multiple action of stress field as in pulsation containers for culturing organisms
Under, promoting iuntercellular to set up and connect and induced growth, endothelium (neural) cell induction around the most celliferous gelatine silk divides
Change and form internal diameter and be about the blood vessel (neural) of 1mm, after gel degradation, ultimately form have spontaneous contractions ability containing blood vessel
With neural atrium.This atrium can reach the purpose of reparative regeneration directly against being attached on disease damage heart.
The preparation of embodiment 2 liver precursor
(1) prepared by material
Extract human bone marrow stem cell (BSCs), fat stem cell (ADSCs), endotheliocyte (ECs), neurocyte regulating liver-QI
Cell, subculture, obtain seed cell；Take 1 Fibrinogen and 7g gelatin is placed in 100mL culture fluid and makes it the most molten
Solving, wherein add the antitumor drug astragalus polysaccharides of 0.1%, sterilization treatment obtains Fibrinogen gelatin astragalus polysaccharides water-setting
Glue material；Taking 4g gelatin to be placed in 100mL culture fluid and make it be completely dissolved, sterilization treatment obtains gelatin solution；Preparation 0.05g/
The thrombin PBS polymer fluid of ml；PU is dissolved in TEG and obtains the synthesis macromolecular solution that mass body volume concentrations is 5%,
Sterilization treatment is stand-by；
The bone marrow stem cell hepatocyte of above-mentioned preparation is mixed with Fibrinogen gelatin astragalus polysaccharides hydrogel material
Obtain fat stem cell and hepatocellular density is 1 × 106The hydrogel containing hepatocyte fat stem cell of individual/mL.Endothelium
Cell, fat stem cell and 0.1% heparin phosphate buffer (PBS) are mixed to get concentration and are 5 × 107The interior sebum of individual/mL
Fat stem cell suspension.Neurocyte and fat stem cell are mixed to get concentration and are 5 × 107The neural fat stem cell of individual/mL
Suspension.
(2) prepared by inner core
Rack surface after sterilization is coated with the gelatin that a layer thickness is about 5mm, reduces temperature and solidify to form to 10～12 DEG C
Inner core, coats hydrogel outside the liver precursor inner core of belt supporting frame and cross-linked polymeric forms one layer of protection insoluble in culture fluid
Film.
(3) shape
By the Fibrinogen gelatin astragalus polysaccharides hydrogel containing hepatocyte bone marrow stem cell and the most celliferous water-setting
Glue is respectively charged into the first squash type shower nozzle 113 and the first squash type shower nozzle 114, endothelium fat stem cell heparin suspension and god
Being respectively charged into the first spray-type shower nozzle 104 and the second spray-type shower nozzle 105 through fat stem cell suspension, thrombin polymer fluid fills
Entering the 4th spray-type shower nozzle 115, synthesis macromolecule loads the 3rd spray-type shower nozzle 108.The support of band gelatin inner core is arranged on
In forming containers 102 and it is connected with motor output shaft, and in forming containers, loads the cell culture fluid of 100mL.Whole device
It is placed in the environment that temperature is 10 DEG C, starts forming device before importing liver after body Model and start to shape.
(4) molten core
After shaping, the liver precursor containing gelatin inner core is placed in the isoperibol of about 37 DEG C, makes gelatin inner core " melt
Change ", take out support, i.e. obtain liver precursor.
(5) later stage cultivates
Liver precursor is placed in incubator cultivation, and in incubation, iuntercellular is set up and is connected and breed growth, without thin
Endothelium (neural) cell and fat stem cell induction around the gelatine silk of born of the same parents are differentiated to form the blood vessel (god that internal diameter is about 1mm
Warp), after gel degradation, ultimately form liver.
The preparation of embodiment 3 bladder precursor
(1) prepared by material
Extract body fat stem cell (ADSC), endotheliocyte (EC), neurocyte and bladder cells, subculture, obtain
To cell material；Take 1g sodium alginate, 1g chitosan and 1g gelatin to be placed in 100mL culture fluid and make it be completely dissolved, at sterilizing
Reason obtains sodium alginate chitosan gelatin hydrogel material；Prepare the calcium chloride water of 0.05g/ml as its cross-linking agent；
Taking 2g glucose and 2g dextrose is placed in 100mL culture fluid and makes it be completely dissolved, it is molten that sterilization treatment obtains dextrose
Liquid；Being dissolved in TEG by PLGA and obtain the solution that volumetric concentration is 5%, sterilization treatment is stand-by；
The fat stem cell bladder cells of above-mentioned preparation is obtained with sodium alginate chitosan gelatin hydrogel material mixing
Density to fat stem cell and bladder cells is 1 × 106The hydrogel containing bladder cells fat stem cell of individual/mL；In
Chrotoplast and fat stem cell are mixed to get concentration and are 5 × 107The endothelium fat stem cell suspension of individual/mL；Neurocyte and
Fat stem cell is mixed to get concentration and is 5 × 107The neurocyte fat stem cell suspension of individual/mL.
(2) prepared by inner core
Shape and structure feature according to bladder inner chamber, by the ellipse chondritic of three-dimensional drawing software design, uses quick shaping method
Make the mould with this inner-cavity structure.Propping up after sterilizing is erected in mould, in mould, adds appropriate gelatin solution,
Reducing temperature makes gelatin solidify to 10～12 DEG C, forms the histoorgan inner core of center band support, at the liver precursor of belt supporting frame
Coat hydrogel outside inner core and cross-linked polymeric forms one layer of protecting film insoluble in culture fluid.
(3) shape
By the sodium alginate chitosan gelatin hydrogel containing bladder cells fat stem cell and the most celliferous hydrogel
Being respectively charged into the first squash type shower nozzle 113 and the first squash type shower nozzle 114, endotheliocyte fat stem cell suspension and nerve are thin
Born of the same parents' fat stem cell suspension is respectively charged into the first spray-type shower nozzle 104 and the second spray-type shower nozzle 105, and calcium chloride cross-linking agent fills
Entering the 4th spray-type shower nozzle 115, synthesis macromolecule loads the 3rd spray-type shower nozzle 108.The support of band gelatin inner core is arranged on
In forming containers 102 and it is connected with motor output shaft, and in forming containers, loads the cell culture fluid of 100mL.Whole device
It is placed in the environment that temperature is 8～10 DEG C, after importing bladder model in a computer, starts forming device, start to shape.
(4) molten core
After shaping, the bladder precursor containing gelatin inner core is placed in calorstat cultivation, removes inner core, take out support,
I.e. obtain the bladder precursor of band inner chamber.
(5) later stage cultivates
Being placed in calorstat by bladder precursor and cultivate, in incubation, iuntercellular is set up and is connected and breed growth,
Endothelium (neural) cell and fat stem cell induction around the most celliferous gelatine silk are differentiated to form blood vessel (neural), at gel
After degraded, ultimately form the bladder with neural and blood vessel.
The preparation of embodiment 4 kidney precursor
(1) prepared by material
Extract blood of human body stem cell, endotheliocyte (EC), neurocyte and nephrocyte, subculture, obtain cell material
Material；Taking 1g sodium alginate and 1g hyaluronic acid is placed in 100mL culture fluid and makes it be completely dissolved, sterilization treatment obtains alginic acid
Sodium hyaluronic acid gel material；Prepare the calcium chloride water of 0.05g/ml as its cross-linking agent；Take 4g gelatin to be placed in
Making it be completely dissolved in 100mL culture fluid, sterilization treatment obtains gelatin solution；Synthesis macromolecule PU is dissolved in TEG
To the synthesis macromolecular solution that mass body volume concentrations is 5%, sterilization treatment is stand-by；
The blood stem cell nephrocyte of above-mentioned preparation and sodium alginate hyaluronic acid gel material mixing are obtained blood
The density of liquid stem cell and nephrocyte is 1 × 106The hydrogel containing nephrocyte blood stem cell of individual/mL.Endotheliocyte and
Blood stem cell is mixed to get concentration and is 4 × 107The endotheliocyte blood stem cell suspension of individual/mL.Neurocyte and blood
Stem cell is mixed to get concentration and is 4 × 107The neurocyte blood stem cell suspension of individual/mL.
(2) prepared by inner core
It is coated with the gelatin of thick layer about 5mm at rack surface, reduces temperature and solidify to form inner core to 10～12 DEG C, at belt supporting frame
Liver precursor inner core outside coat hydrogel cross-linked polymeric and form one layer of protecting film insoluble in culture fluid.
(3) shape
Hydrogel containing nephrocyte blood stem cell and the most celliferous hydrogel are respectively charged into the first squash type shower nozzle
113 and the second squash type shower nozzle 114, endotheliocyte blood stem cell solution and neurocyte blood stem cell solution fill respectively
Entering the first spray-type shower nozzle 104 and the second spray-type shower nozzle 105, calcium chloride cross-linking agent/polymer fluid loads the 4th spray-type shower nozzle
115, synthesis macromolecule loads the 3rd spray-type shower nozzle 108.In the support of band gelatin inner core is arranged on forming containers 102 and with
Motor output shaft is connected, and loads the cell culture fluid of 100mL in forming containers.It is 8～10 DEG C that whole device is placed in temperature
Environment in, import and start forming device after kidney model, start to rotate stack shaping.
(4) molten core
After shaping, the kidney precursor containing gelatin film inner core is placed in calorstat cultivation, makes gelatin film inner core
" melt ", take out support, i.e. obtain kidney precursor.
(5) later stage cultivates
Kidney precursor is placed in incubator cultivation, and in incubation, iuntercellular is set up and is connected and breed growth, without thin
Endothelium (neural) cell and blood stem cell induction around the gelatine silk of born of the same parents are differentiated to form the blood vessel (god that internal diameter is about 1mm
Warp), after gel degradation, ultimately form and there is blood vessel and neural kidney.
The preparation of embodiment 5 breast prosthesis
(1) prepared by material
Extract body fat stem cell (ADSC), endotheliocyte (EC), neurocyte and lymphocyte, subculture, obtain
To cell material；Taking 1g sodium alginate and 2g gelatin is placed in 100mL culture fluid and makes it be completely dissolved, sterilization treatment obtains Sargassum
Acid sodium gelatin hydrogel material；Prepare the calcium chloride water of 0.05g/ml as its cross-linking agent/polymerization；Take 2g gelatin to be placed in
Making it be completely dissolved in 100mL culture fluid, sterilization treatment obtains gelatin solution；Synthesis macromolecule PU is dissolved in TEG
To the synthesis macromolecular solution that mass body volume concentrations is 5%, sterilization treatment is stand-by；
The fat stem cell of above-mentioned preparation and sodium alginate gelatin hydrogel material mixing are obtained fat stem cells density
It is 1 × 106The hydrogel of the fatty stem cell of individual/mL.Wherein adding 1 μM of insulin, 1 μM of dexamethasone, 50u/mL presses down
Peptidase and 0.5mmol/L isobutyl methyl xanthine derivative.Neurocyte and fat stem cell are mixed to get concentration and are 3
×108The neurocyte fat stem cell suspension of individual/mL.Fat stem cell suspension adds 50ng/mL endothelial cell growth factor (ECGF)
(VEGF), 3ng/mL TGF β 1 somatomedin, 10ng/mL fibroblast growth factor (b FGF).
(2) prepared by inner core
It is coated with the gelatin of thick layer about 5mm at rack surface, reduces temperature and solidify to form inner core to 10～12 DEG C, at belt supporting frame
Liver precursor inner core outside coat hydrogel cross-linked polymeric and form one layer of protecting film insoluble in culture fluid.
(3) shape
By fatty stem cell, insulin, dexamethasone, aprotinin and the Sargassum of isobutyl methyl xanthine derivative
Acid sodium gelatin hydrogel and the most celliferous hydrogel are respectively charged into the first squash type shower nozzle 113 and the second squash type shower nozzle
114, endotheliocyte fat stem cell suspension and neurocyte fat stem cell suspension are respectively charged into the first spray-type shower nozzle 104
With the second spray-type shower nozzle 105, calcium chloride cross-linking agent loads the 4th spray-type shower nozzle 115, and synthesis macromolecule loads the 3rd sprinkling
Formula shower nozzle 108.In the support of band gelatin inner core is arranged on forming containers 102 and it is connected with motor output shaft, and describes at one-tenth
The cell culture fluid of 100mL is loaded in device.Whole device is placed in the environment that temperature is 10～12 DEG C, opens after importing breast model
Dynamic forming device, starts to shape.
(4) molten core
After shaping, the breast precursor containing gelatin film inner core is placed in calorstat cultivation, makes gelatin inner core " melt
Change ", take out support, i.e. obtain breast precursor.
(5) later stage cultivates
In incubation, iuntercellular is set up and is connected and breed growth, the endothelium (god around the most celliferous gelatine silk
Warp) cell and fat stem cell induction be differentiated to form the blood vessel (neural) that internal diameter is about 1mm, after gel degradation, ultimately form
There is the breast of certain function.
Claims (6)
1. one kind utilizes the method that rotation method of piling prepares histoorgan, it is characterised in that the method uses following special equipment
Being prepared, described equipment includes support means, many nozzle components, motion, forming device and control system；Described
Support arrangement includes supporting leg and base plate (101)；Described motion uses guide rail slide block mechanism, including X direction guiding rail (121), X
Slide to slide block (120), Y-direction guide rail (107), the first Y-direction slide block (111), the second Y-direction slide block (112), Z-direction guide rail (118), Z-direction
Block (106)；Described X direction guiding rail (121) is located on base plate (101), and Z-direction guide rail (118) is fixed in X on slide block (120), Y-direction
Guide rail (107) is fixed on Z-direction slide block (106)；Described many nozzle components at least contain six shower nozzles, divide two groups and are separately mounted to
On first Y-direction slide block and the second Y-direction slide block；Described forming device includes forming containers (102), support (103) and motor
(119), forming containers (102) is arranged on base plate (101) and is in the lower section of nozzle component, and support (103) is arranged on shaping
In container (102) and it is connected with motor (119) output shaft；
Its preparation method comprises the following steps:
1) prepared by material:
Preparation 0.2～20% (w/v) hydrogel solution, 1～10% (w/v) cross-linking agent/polymer fluid, 0.1～10% (w/v) synthesis
Macromolecular solution, 0.2～10% (w/v) gelatin solution and cell culture fluid；Separating or induction seed cell, seed cell is close
Degree is 1 × 103～1 × 106Individual/mL, is mixed seed cell and hydrogel solution containing seed by 1～9:9～1 volume ratio
The hydrogel solution of cell；Separate or inducing endothelial cell and neurocyte make cell suspension, endotheliocyte suspension and
Neurocyte suspension density is 1 × 105～1 × 108Individual/mL；
2) prepared by inner core
When preparation has the histoorgan of inner chamber, according to the cavity shape of histoorgan to be prepared, use three-dimensional drawing software
Design its threedimensional model, make the mould with this inner-cavity structure, when the histoorgan of forming solid, mould by quick shaping method
Tool is the cylinder of diameter thick 8～10mm；Support (103) is put into mould central authorities, after adding gelatin solution in mould, reduces temperature
Degree makes gelatin solidify to 8～10 DEG C, opens mould and obtains the histoorgan inner core (401) of center band support (103), at belt supporting frame
Histoorgan inner core (401) coat outward one layer of hydrogel solution, and with cross-linking agent/polymer fluid crosslinking/polymerization；
3) shape
Hydrogel solution containing seed cell and the most celliferous hydrogel solution are respectively charged into the first squash type shower nozzle (113)
With in the second squash type shower nozzle (114), cross-linking agent/polymer fluid, neurocyte suspension, endotheliocyte suspension and synthesis macromolecule
Solution is respectively charged into the 4th spray-type shower nozzle (115), the second spray-type shower nozzle (105), the first spray-type shower nozzle (104) and the 3rd
In spray-type shower nozzle (108), in forming containers, load cell culture fluid (405), support (103) is arranged on forming containers
(102) in and it is connected with motor (119) output shaft, after the model of histoorgan to be formed is imported control system, starts and shape
Equipment, starts stack shaping；In forming process, support (103) rotates around Y negative sense, and shower nozzle does translational motion, treats as
During the main part of shape histoorgan, the first squash type shower nozzle (113) moves to the water-setting specifying position extrusion containing seed cell
Glue is wrapped on inner core, and the 4th spray-type shower nozzle (115) equipped with cross-linking agent/polymer fluid moves to the first squash type shower nozzle simultaneously
(113) dead astern sprays cross-linking agent/polymer fluid and cross-links it/be polymerized；When shaping blood vessel, the second squash type shower nozzle
(114) move to specify position to extrude the most celliferous hydrogel be wrapped on inner core, the 4th spray-type shower nozzle (115) fortune simultaneously
Moving the second squash type shower nozzle (114) dead astern, the first spray-type shower nozzle (104) and the 3rd spray-type shower nozzle (108) are transported respectively
Moving the second squash type shower nozzle (114) dead ahead, under the inner core rotated drives, the new hydrogel being wound around moves to the 4th successively
Carry out cross-linking/being polymerized under spray-type shower nozzle (115), wash through culture fluid, to the first spray-type shower nozzle (104), the 3rd
Carry out under spray-type shower nozzle (108) spraying endotheliocyte and composite synthesis macromolecular solution；As body and the spirit through time, the second squash type
Shower nozzle (114) moves to specify position to extrude the most celliferous hydrogel and is wrapped on inner core, the 4th spray-type shower nozzle simultaneously
(115) the second squash type shower nozzle (114) dead astern, the second spray-type shower nozzle (105) and the 3rd spray-type shower nozzle (108) are moved to
Moving to the second squash type shower nozzle (114) dead ahead respectively, under the inner core rotated drives, the new hydrogel being wound around moves successively
Carry out cross-linking/being polymerized to the 4th spray-type shower nozzle (115), wash through culture fluid, to the second spray-type shower nozzle
(105), carry out under the 3rd spray-type shower nozzle (108) spraying neurocyte and composite synthesis macromolecule；So repeatedly it is wound around accumulation
Until forming the histoorgan precursor containing inner core；
4) subsequent treatment
Histoorgan precursor containing inner core is placed in calorstat cultivation, molten except taking out support after inner core, when continuing to cultivate one section
Forming revolution shape histoorgan after between, nerve and blood vessel (502) are the most therethrough.
A kind of utilization the most as claimed in claim 1 rotates the method that method of piling prepares histoorgan, it is characterised in that: described
Frame is that two ends are cylindrical, middle for prismatic structure.
A kind of utilization the most as claimed in claim 2 rotates the method that method of piling prepares histoorgan, it is characterised in that described
The material of frame is rustless steel or titanium alloy.
A kind of utilization the most as claimed in claim 1 rotates the method for piling method of preparing histoorgan, it is characterised in that: described the
The little guide rail (110) and with the first of Y-direction guide rail parallel is had on one Y-direction slide block (111) and the second Y-direction slide block (112)
It is respectively mounted the first small slide block (109) and the second small slide block on two little guide rails (117), the first little guide rail and the second little guide rail
(116), the first small slide block and the second small slide block have the hole of stationary nozzle.
A kind of utilization the most as claimed in claim 1 rotates the method that method of piling prepares histoorgan, it is characterised in that: described many
Nozzle component includes at least two squash type shower nozzle and four spray-type shower nozzles, and the nozzle inside diameter of two squash type shower nozzles is respectively
200 μm～1mm and 50 μm～500 μm, the mesh diameter of spray-type spray nozzle is 50 μm～100 μm.
A kind of utilization the most as claimed in claim 1 rotates the method that method of piling prepares histoorgan, it is characterised in that: described
Seed cell is to have the stem cell of differentiation capability or have the adult cell of physiologically active, and described stem cell is that fat is dry thin
Born of the same parents, embryonic stem cell, blood stem cell, bone marrow stem cell or induction type versatile stem cell, described adult cell is that fat is thin
Born of the same parents, hepatocyte, bladder cells, lymphocyte, myocardial cell or nephrocyte；Described hydrogel is for being compounded with gelatin, collagen, substrate
The sodium alginate of one or more in laminin in glue, carrageenan, chitosan, agar, hyaluronic acid, matrigel, elastin laminin
Or fibrinogen solution；Compound various cell cryopreservation agent, cell growth factor, medicine, anticoagulant in described hydrogel；
Cell cryopreservation agent is one or more complex in dimethyl sulfoxide, glycerol, dextrose；Described cell growth factor is blood
Endothelial tube somatomedin, basic fibroblast growth factor, hepatocyte growth factor, human blood platelets derived growth factor, turn
Change one or more complex in growth factor beta 1；Described medicine is antitumor drug, and this antitumor drug is that the Radix Astragali is many
One in diamino dichloro network platinum sugared, cis, mitomycin, 5-fluorouracil, Claritin, antibiotic and viral vaccine
Or multiple complex；Described anticoagulin is one or both complex in heparin and paclitaxel；Described synthesis high score
The solute of sub-solution be polyurethane, polycaprolactone, Merlon, Polyethylene Glycol, Poly(D,L-lactide-co-glycolide, polyester and
One or more in polyhydroxy acid ester, solvent is TEG or Isosorbide-5-Nitrae-dioxane.
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| CN201410473055.4A CN104207859B (en) | 2014-09-16 | 2014-09-16 | Rotation method of piling is utilized to prepare method and the special equipment of histoorgan |
| PCT/CN2014/089769 WO2016041238A1 (en) | 2014-09-16 | 2014-10-29 | Method and dedicated device for preparing tissue and organ by using spin accumulation method |
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| CN201410473055.4A CN104207859B (en) | 2014-09-16 | 2014-09-16 | Rotation method of piling is utilized to prepare method and the special equipment of histoorgan |
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| CN114456929A (en) * | 2021-11-25 | 2022-05-10 | 浙江大学 | Device and method for constructing localized cell load model in three-dimensional scaffold |
| CN114789046B (en) * | 2022-05-23 | 2024-04-02 | 燕山大学 | Heavy metal trapping agent and application thereof |
| CN116285382A (en) * | 2023-02-15 | 2023-06-23 | 华中科技大学同济医学院附属同济医院 | 3D printing artificial uterus and preparation method and application thereof |
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| US7815763B2 (en) * | 2001-09-28 | 2010-10-19 | Abbott Laboratories Vascular Enterprises Limited | Porous membranes for medical implants and methods of manufacture |
| US8431060B2 (en) * | 2006-01-31 | 2013-04-30 | Abbott Cardiovascular Systems Inc. | Method of fabricating an implantable medical device using gel extrusion and charge induced orientation |
| CN101692987B (en) * | 2009-10-16 | 2011-04-27 | 清华大学 | Rotating disc type multi-nozzle three-dimensional controlled forming system for complex organ precursor |
| US20120089238A1 (en) * | 2010-10-06 | 2012-04-12 | Hyun-Wook Kang | Integrated organ and tissue printing methods, system and apparatus |
| CN102319126A (en) * | 2011-07-21 | 2012-01-18 | 清华大学 | Fixed multi-nozzle complex organ precursor three-dimensional controlled forming system |
| CN102599990B (en) * | 2012-03-23 | 2015-04-15 | 清华大学 | Composite type complex organ precursor three-dimensional controlled forming system with multiple sprayers |
| CN102755203B (en) * | 2012-07-13 | 2015-01-21 | 清华大学 | Spraying and coating combined complex tissue organ manufacturing system |
| CN203263583U (en) * | 2013-05-03 | 2013-11-06 | 清华大学 | Multi-freedom-degree pneumatic multi-spraying-nozzle manufacturing system for complex tissue and organs |
| CN103462725B (en) * | 2013-08-06 | 2015-08-26 | 浙江大学 | A kind of three-dimensional biological structure printing equipment and method |
| CN103919629B (en) * | 2014-04-18 | 2016-09-28 | 清华大学 | A kind of toughness organizational structure and 3D printing-forming equipment thereof and method |
| CN103948456B (en) * | 2014-04-22 | 2016-01-13 | 上海大学 | The pneumatic many shower nozzles of the rotating disc type biological 3D printing-forming system and method for Automated condtrol |
| CN204072388U (en) * | 2014-09-16 | 2015-01-07 | 清华大学 | A kind of special equipment utilizing rotation method of piling to prepare histoorgan |
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2014
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| WO2016041238A1 (en) | 2016-03-24 |
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