CN106361463A - Liver tissue simulation structure and forming method thereof - Google Patents
Liver tissue simulation structure and forming method thereof Download PDFInfo
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- CN106361463A CN106361463A CN201510438007.6A CN201510438007A CN106361463A CN 106361463 A CN106361463 A CN 106361463A CN 201510438007 A CN201510438007 A CN 201510438007A CN 106361463 A CN106361463 A CN 106361463A
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Abstract
The invention discloses a liver tissue simulation structure and a forming method thereof, and belongs to the technical field of organ manufacturing, bio-materials and medical appliances. The structure comprises a main body structure, an artery blood vessel system, a vein blood vessel system, a nerve system and a bile duct system. The blood vessel systems, the nerve system and part of the bile duct system are distributed in the main body structure in a branch shape. The liver tissue simulation structure prepared by utilizing upper moulds of different diameters, multi-step lower moulds and inner moulds which simulates the artery/vein, nerve and bile duct systems ensures organic arrangement of the artery, vein, nerve and bile duct systems of different directions in the same structure body, is closer to a true morphology of human organs, and can finish various physiological functions of the liver. The method is simple and convenient in forming and low in cost.
Description
Technical field
The invention belongs to the artificial manufacturing technology field of organism complicated tissue organ, relate to the use of cell, synthesis macromolecular material,
Cell matrix materials prepare the process of complex organization's artificial liver, organizational project technique different from the past.
Background technology
The proposition of tissue engineering is artificial organ and the manufacture of organ and culture specifies a way forward.Especially in the present
In the case that its organ transplantation technique is more and more ripe, and corresponding organ origin is more and more in short supply, develop artificial organ organ
Manufacture and just become the task of top priority promoting the well-being of mankind.
Although artificial organ organ transplantation or a relatively jejune field, need long-term accumulation and exploration, and be related to it
His ethics morals problem, but had been developed that the relatively simple artificial tissue of the structures such as skeleton, cartilage, skin at present,
And have been obtained for certain clinical practice.The present invention, on the basis of previous work, by constantly putting into practice, finds out a set of
The manufacture method of new artificial liver, makes complicated device pipe, such as liver, heart, kidney, breast, manufacture significantly stride forward
One step.
Content of the invention
It is an object of the invention to provide a kind of liver organization model configuration and its manufacturing process are so as to simulate sky in function and structure
Right animal body liver, provides new probability for fields such as organ manufacture, transplanting.
Technical scheme is as follows:
A kind of liver organization model configuration it is characterised in that: described liver organization model configuration includes agent structure, arteries
System, venous vasculature, nervous system and biliary system；
Described arterial vascular system, venous vasculature and nervous system are arranged on inside agent structure, and part biliary system is divided
In agent structure, another part is distributed in outside agent structure cloth；It is careful that described agent structure is followed successively by kind containing blood vessel from the inside to the outside
The hydrogel layer of born of the same parents, the hydrogel layer of seed cell containing liver, the hydrogel layer containing stem cell and protective layer；
Described arterial vascular system and venous vasculature are branch's tubular structure that the hydrogel containing blood vessel seed cell is constituted；Institute
State the branched fiber binding structure that nervous system (103) is that the hydrogel containing neural seed cell is constituted, described biliary system is containing gallbladder
The conduit of pipe epithelial cell；Biliary system and vascular system are spaced in agent structure.
The outline of agent structure of the present invention is in shape, spindle shape, spheroid form or the ellipsoid shape of animal body liver.
Described protective layer is synthesis macromolecular material.
Sodium alginate that hydrogel of the present invention is all 0.1～20% using mass percentage concentration, bioactive peptide, collagen, heparin,
Chondroitin sulfate, hyaluronic acid, mucopolysaccharide, glycoprotein, matrigel, dextrose, shitosan, gelatin and Fibrinogen are molten
At least one in liquid.Described liver seed cell is hepatocyte, or stem cell regulating liver-QI cell mixture, or stem cell and growth
Factor cocktails；Stem cell is at least one in fat stem cell, embryonic stem cell and induced multi-potent stem cell；Described blood vessel
Seed cell is fat stem cell, mesenchymal stem cells MSCs, vascular endothelial cell, smooth muscle cell, fibroblast, embryo
At least one in tire stem cell and induced multi-potent stem cell；Described nerve seed cell is neuron, or Schwann cell, or
Stem cell and growth factor mixture.Described synthesis macromolecular material adopts polyethylene, polycaprolactone, polyurethane, polytetrafluoro
Ethylene, the complex of polylactic acid, polyester and one or more of lactic acid and ethanol copolymer material.
The present invention provide a kind of liver organization model configuration preparation method it is characterised in that: the method comprises the steps:
1) prepare particular manufacturing craft: prepare described particular manufacturing craft using machining or the method for 3 D-printing, this mould include lower mould,
Inner mold, simulation biliary system mould and mold；Described inner mold include simulate arterial vascular system inner mold, simulation quiet
Arteries and veins vascular system inner mold and the inner mold of simulated nervous system；
2) three inner molds of simulation tremulous pulse, vein and nervous system are inserted and fixed by three holes at first order mold top
Good, then it is positioned on the first order step of lower mould fixing together；Simulation biliary system mould is passed through the hole on lower mould
Fixing；Afterwards the hydrogel containing blood vessel seed cell is filled in the gap between inner mold and first order mold, and adopts
Cross-linking agent makes hydrogel containing blood vessel seed cell crosslinked, removes first order mold, formed stable containing blood vessel seed cell
Hydrogel layer；
3) it is placed on using the diameter second level mold bigger than the first order on the second level step of lower mould fixing, liver kind will be contained
The hydrogel of daughter cell is filled in the gap between the hydrogel layer containing blood vessel seed cell and second level mold, and using friendship
Connection agent makes the hydrogel containing liver seed cell crosslinked, removes second level mold, forms the stable water-setting of seed cell containing liver
Glue-line；
4) according to step 3) method, be placed on the of lower mould using the diameter third level mold bigger than second level mold
Fixing on three-level step, the hydrogel containing stem cell is filled between liver seed cell hydrogel layer and third level mold
In gap, and make the hydrogel containing stem cell crosslinked using cross-linking agent, obtain the hydrogel layer containing stem cell；
5) remove inner mold and lower mould, irrigate respectively in the gap that simulation tremulous pulse and simulation venous vasculature inner mold stay
Vascular endothelial cell, forms arterial vascular system and simulation venous vasculature, the gap staying in simulated nervous system inner mold
Middle perfusion Schwann cell suspension, forms branch's nervous system；
6) exposed part biliary system outside agent structure is contained hepatocellular hydrogel parcel with one layer, and made using cross-linking agent
Celliferous hydrogel is crosslinked；
7) synthesis macromolecular material is dissolved in and in organic solvent, makes the synthesis macromolecular solution that mass percentage concentration is 5%～30%,
It is placed in spray bottle, become macromolecular material in agent structure surface even application one lamination shaping, be then placed in pbs molten
Extract in liquid, finally give stable liver organization model configuration.
In the method for the present invention, described mold, lower mould, simulation arterial vascular system inner mold, simulation venous vasculature
Inner mold, simulated nervous system inner mold and simulation biliary system inner mold adopt metal or synthesis macromolecular material；Described metal
Using aluminium alloy, pyrite or stainless steel material；Synthesis macromolecular material adopt politef, pbs plastics, photosensitive resin,
Rubber, acrylonitrile-butadiene-styrene copolymer plastics, polystyrene plastics, poly methyl methacrylate plastic, polyformaldehyde
Plastics, polypropylene plastics, vinyon, polrvinyl chloride, Maranyl, polycarbonate plastic, epoxy resin, poly- oneself
Lactone, polyurethane or silicone plasties.
The present invention compared with prior art, have the advantage that and salience technique effect: 1. the present invention liver organization simulation knot
Structure, has tremulous pulse, vein, nerve and bile duct tissue's network, any artificial organ is all complicated than ever, closer to human organ
Real topography, and the various physiological functions of liver can be completed.2. the manufacturing process of the present invention ensure that different trends tremulous pulsies,
The organic arrangement in same structure body of vein, nerve and biliary system.3. the method is simple for production, low cost.
Brief description
Fig. 1 is liver organization model configuration schematic diagram.
Fig. 2 is the mould structure schematic diagram preparing liver organization model configuration.
In figure: 101- arterial vascular system；102- venous vasculature；103- nervous system；104- protective layer；105- contains dry thin
The hydrogel layer of born of the same parents；106- seed cell containing liver hydrogel layer；107- contains the hydrogel layer of blood vessel seed cell；108- Within Human Biliary Tract
System.
Mould under 201-；202a- simulates arterial vascular system inner mold；202b- simulates venous vasculature inner mold；202c-
Simulated nervous system inner mold；203 first order molds；204- second level mold；205- simulates biliary system inner mold.
Specific embodiment
The present invention is described further below in conjunction with the accompanying drawings.
As shown in figure 1, a kind of liver organization model configuration that the present invention provides includes agent structure, arterial vascular system 101, quiet
Arteries and veins vascular system 102, nervous system 103 and biliary system 108；Described arterial vascular system 101, venous vasculature
102 and nervous system 103 be arranged on inside agent structure, part biliary system 108 is distributed in agent structure, another part
It is distributed in outside agent structure；Described agent structure outline is in shape, spindle shape, spheroid form or the ellipsoid of animal body liver
Shape.
Described agent structure is followed successively by hydrogel layer 107 containing blood vessel seed cell, the hydrogel layer of seed cell containing liver from the inside to the outside
106th, hydrogel layer 105 and the protective layer 104 of stem cell are contained；Described arterial vascular system and venous vasculature are kind containing blood vessel
Branch's tubular structure that the hydrogel of daughter cell is constituted；Described nervous system 103 is that the hydrogel containing neural seed cell is constituted
Branched fiber binding structure, described biliary system is the conduit containing bile duct epithelial cell；Biliary system is spaced at vascular system
In agent structure.
Sodium alginate that described hydrogel is all 0.1～20% using mass percentage concentration, bioactive peptide, collagen, heparin, sulphuric acid are soft
In ossein, hyaluronic acid, mucopolysaccharide, glycoprotein, matrigel, dextrose, shitosan, gelatin and fibrinogen solution
At least one.
Described liver seed cell is hepatocyte, or stem cell regulating liver-QI cell mixture, or stem cell and growth factor mixture；Dry
Cell is at least one in fat stem cell, embryonic stem cell and induced multi-potent stem cell；Described blood vessel seed cell is fat
Stem cell, mesenchymal stem cells MSCs, vascular endothelial cell, smooth muscle cell, fibroblast, embryonic stem cell and induction
At least one in pluripotent stem cell；Described nerve seed cell be neuron, or Schwann cell, or stem cell with growth because
Sub- mixture.
Described protective layer 104 be synthesis macromolecular material, synthesis macromolecular material adopt polyethylene, polycaprolactone, polyurethane,
Politef, the complex of polylactic acid, polyester and one or more of lactic acid and ethanol copolymer material.
A kind of preparation method of liver organization model configuration that the present invention provides, the method comprises the steps:
1) prepare particular manufacturing craft: the method using machining or 3 D-printing prepares described particular manufacturing craft, and this mould includes lower mould
201st, inner mold, simulation biliary system mould and mold；Described inner mold includes simulating arterial vascular system inner mold, mould
Intend the inner mold of venous vasculature inner mold and simulated nervous system；Mold, the material of lower mould, inner mold and outer mold
Can be using metal or synthesis macromolecular material, such as aluminium alloy, pyrite, rustless steel, politef, acrylonitrile-butadiene-benzene
Ethylene copolymer (abs) plastics, photosensitive resin, rubber, polystyrene (ps) plastics, polymethyl methacrylate (pmma)
Plastics (lucite), polyformaldehyde (pom) plastics, polypropylene (pp) plastics, polyethylene (pe) plastics, polrvinyl chloride
(pvc), polyamide (pa) plastics (nylon), polycaprolactone (pcl), polyurethane (pu), Merlon (pc) plastics,
Epoxy resin (ep) or silicone plasties (is).
Lower mould inside is to become big stepped from the inside to the outside successively, depending on the substrate number of plies that the series of step can make as needed.
The material of lower mould synthesizes macromolecular material for hard, as shown in Figure 2 () taking level Four step as a example.
The mold of different-diameter is corresponding with the step of same diameter in lower mould.The outer casing thickness of the mold of different-diameter with
In lower mould, step width at different levels are corresponding.
2) three holes by first order mold 203 top for three inner molds 202 of tremulous pulse, vein and nervous system will be simulated
It is inserted and fixed, be then positioned on the first order step of lower mould 201 fixing together；Biliary system mould 205 will be simulated
Fix through the hole on lower mould 201；Afterwards the hydrogel containing blood vessel seed cell is filled into inner mold 202 and the first order
In gap between mold 203, and make the hydrogel containing blood vessel seed cell crosslinked using cross-linking agent, remove mould in the first order
Tool 203, forms the stable hydrogel layer 107 containing blood vessel seed cell；
3) it is placed on the second level step of lower mould 201 fixing using the diameter second level mold 204 bigger than the first order, will
Hydrogel containing liver seed cell is filled between hydrogel layer 107 and the second level mold 204 containing blood vessel seed cell
In gap, and make the hydrogel containing liver seed cell crosslinked using cross-linking agent, remove second level mold 204, formed stable
The hydrogel layer of seed cell containing liver 106；
4) according to step 3) method, be placed on lower mould 201 using the diameter third level mold bigger than second level mold
Third level step on fixing, by the hydrogel containing stem cell be filled into liver seed cell hydrogel layer and third level mold it
Between gap in, and make hydrogel containing stem cell crosslinked using cross-linking agent, obtain the hydrogel layer containing stem cell；
5) remove inner mold and lower mould, irrigate respectively in the gap that simulation tremulous pulse and simulation venous vasculature inner mold stay
Vascular endothelial cell, forms arterial vascular system and simulation venous vasculature, the gap staying in simulated nervous system inner mold
Middle perfusion Schwann cell suspension, forms branch's nervous system；
6) exposed part biliary system outside agent structure is contained hepatocellular hydrogel parcel with one layer, and made using cross-linking agent
Celliferous hydrogel is crosslinked；
7) synthesis macromolecular material is dissolved in and in organic solvent, makes the synthesis macromolecular solution that mass percentage concentration is 5%～30%,
It is placed in spray bottle, become macromolecular material in agent structure surface even application one lamination shaping, be then placed in pbs molten
Extract in liquid, finally give stable liver organization model configuration.
Enumerate several specific embodiments below, to further understand the present invention.
Embodiment 1
1) use politef system with the standby mold containing two-stage step of mach mode, with the standby branch containing one-level of polyurethane
Simulation arterial vascular system, simulation venous vasculature and simulated nervous system inner mold, with polyethylene prepare with micropore
Simulation biliary system inner mold containing one-level branch；Prepare 1% fibrinogen solution, tremulous pulse, vein, nerve and gallbladder will be simulated
Four inner molds of guard system are fixed by three holes at first order mold top and the hole insertion of lower mould respectively, then together
Fixing on the first order step of lower mould.Injection Fibrinogen and vascular endothelial cell between having on inner mold and the first order
Mixture, cell density is 1 × 107Individual/ml, is subsequently adding thrombin solution (20iu/ml) immersion molding and gathers for 2 minutes
Close, remove first order mold, form the stable hydrogel layer containing vascular endothelial cell；
2) it is enclosed within the second level step of base mould using the diameter second level mold bigger than the first order.By Fibrinogen/
(cell density is 1 × 10 to hepatocyte mixture5Individual/ml), it is filled on the hydrogel layer and the second level containing vascular endothelial cell
In gap between mould, being subsequently adding thrombin solution (20iu/ml) makes to gather containing Fibrinogen in hepatocellular hydrogel
Close, remove mold, formed stable containing hepatocellular hydrogel layer；
3) it is enclosed within the third level step of base mould using the diameter third level mold bigger than the second level.By Fibrinogen/
(cell density is 1 × 10 to fat stem cell mixture4Individual/ml), it is filled into containing mould on hepatocellular hydrogel layer and the third level
In gap between tool, being subsequently adding thrombin solution (20iu/ml) makes Fibrinogen polymerization in hydrogel containing hepatocyte,
Remove mold, form the hydrogel layer of stable fatty stem cell；
4) inner mold of simulation arterial vascular system, simulation venous vasculature and simulated nervous system and lower mould are removed, in mould
Intend difference Perfused vessel endotheliocyte in the gap that tremulous pulse and simulation venous vasculature inner mold stay, form arterial vascular system
With simulation venous vasculature, irrigate Schwann cell suspension in the gap that simulated nervous system inner mold stays, formed and divide
Prop up nervous system；
5) exposed part biliary system outside agent structure is contained hepatocellular hydrogel parcel with one layer, and made using cross-linking agent
Celliferous hydrogel is crosslinked；
6) synthesis macromolecule polyurethane is dissolved in and in organic solvent, makes the synthesis macromolecular solution that mass percentage concentration is 5%, and
It is placed in spray bottle, in the agent structure surface even application one strata urethane shaping, be then placed in extraction in pbs solution,
Obtain stable liver organization model configuration eventually.
Embodiment 2:
1) with abs to be raw material prepared in the way of 3d printing containing three-level step lower mould, mold and inner mold, with poly-
The inner mold containing two grades of branches for the ethylene preparation；
2) prepare 1% sodium alginate soln, by three inner molds of simulated blood vessel, nerve and biliary system and outer mold by
Three holes at first order inner mold top are inserted and fixed, and are then positioned on the first order step of lower mould fixing together, and
Will be fixing among the array hole of lower for branch's insertion of inner mold mould.Between inner mold and first order mold, injection contains 1% paclitaxel
Sodium alginate endothelial cell growth factor (ECGF) (egf10ng/ml)/fat stem cell mixture (cell density be 1 × 103Individual/ml),
Being subsequently adding the calcium chloride that w/v is 1% makes sodium alginate cross-linking for 2 minutes, removes first order mold, is formed stable
Hydrogel layer containing endothelial cell growth factor (ECGF)/fat stem cell；
3) it is enclosed within the second level step of lower mould using the diameter second level mold bigger than the first order.Will be thin for sodium alginate/liver
(cell density is 1 × 10 to the intracellular growth factor (hgf0.5ng/ml)/fat stem cell mixture4Individual/ml) it is filled into containing fat
In gap between the hydrogel layer of fat stem cell and second level mold, it is subsequently adding 2 points of the calcium chloride that w/v is 1%
Clock makes sodium alginate cross-linking, removes mold, forms the stable hydrogel layer containing hepatocyte growth factor/fat stem cell；
4) it is enclosed within the third level step of lower mould using the diameter third level mold bigger than the second level.By sodium alginate/fat
Stem cell compositions, are filled into the gap between the hydrogel layer containing hepatocyte growth factor/fat stem cell and third level mold
In, being subsequently adding the calcium chloride that w/v is 1% makes sodium alginate cross-linking for 2 minutes, removes mold, forms stable containing
The hydrogel layer of fat stem cell；
6) remove upper and lower, middle mould, in the gap that simulated nervous system inner mold stays, irrigate Schwann cell suspension, shape
Become branch's nervous system；
7) it is equipped with plga/ TEG (tetraglycol) solution that concentration is 10% (w/v), add 1% (w/w)
Paclitaxel, in spray injection bottle, shape one layer of plga of artificial liver surface even application, be then placed in pbs solution
Middle extraction, obtains final stable liver organization model configuration.
Embodiment 3:
1) the use of degradable metal magnesium is raw material standby mold containing two-stage step in the way of three-dimensional (3d) prints, bed die has,
Inner mold and outer mold；Prepare 0.1% collagen solution, by simulation tremulous pulse, vein, nerve and biliary system four inner molds by
Hole on lower mould is fixed.Injection collagen solution, bone marrow stem cell and endotheliocyte life between inner mold and first order mold
The long factor (egf0.5ng/ml) mixture, wherein bone marrow stem cell density are 1 × 103Individual/ml, is subsequently adding 0.01m's
Sodium hydroxide solution makes collagen polymerization shape for 2 minutes, removes first order mold, formed stable containing bone marrow stem cell and endothelium
The hydrogel layer of cell；
2) it is enclosed within the second level step of base mould using the diameter second level mold bigger than the first order.By Fibrinogen/
(cell density is 1 × 10 to bone marrow stem cell mixture3Individual/ml), add hepatocyte growth factor (hgf0.5ng/ml), people
Platelet derived growth factor (bb or pdgf-bb 50ng/ml), transforminggrowthfactor-β1 (tgf β 1 10ng/ml) and
Basic fibroblast growth factor (b-fgf 205ng/ml), is filled into the hydrogel layer containing bone marrow stem cell and endotheliocyte
In gap and the mold of the second level between, being subsequently adding thrombin solution (20iu/ml) makes the fiber in cellular matrix solution
Fibrinogen polymerization, removes mold and inner mold, forms the hydrogel layer stably containing bone marrow stem cell and somatomedin；
3) remove upper and lower, middle mould, in the gap that simulated nervous system inner mold stays, irrigate Schwann cell suspension, shape
Become branch's nervous system；
4) it is equipped with pcl/ TEG (tetraglycol) solution that concentration is 20% (w/v), add 1% (w/w's)
Heparin, in spray injection bottle, in the one layer of pcl of artificial liver surface even application shaping, is then placed in extraction in pbs solution,
Obtain final stable liver organization model configuration.
Claims (8)
1. a kind of liver organization model configuration it is characterised in that: described liver organization model configuration includes agent structure, arterial blood
Guard system (101), venous vasculature (102), nervous system (103) and biliary system (108)；
Described arterial vascular system (101), venous vasculature (102) and nervous system (103) are arranged on agent structure
Inside, part biliary system (108) is distributed in agent structure, and another part is distributed in outside agent structure；Described agent structure
It is followed successively by the hydrogel layer containing blood vessel seed cell (107), the hydrogel layer of seed cell containing liver (106) from the inside to the outside, contain
The hydrogel layer (105) of stem cell and protective layer (104)；
Described arterial vascular system and venous vasculature are branch's tubular structure that the hydrogel containing blood vessel seed cell is constituted；Institute
State the branched fiber binding structure that nervous system (103) is that the hydrogel containing neural seed cell is constituted, described biliary system is containing gallbladder
The conduit of pipe epithelial cell；Biliary system and vascular system are spaced in agent structure.
2. as claimed in claim 1 a kind of liver organization model configuration it is characterised in that: described agent structure outline is in dynamic
The shape of object liver, spindle shape, spheroid form or ellipsoid shape.
3. as claimed in claim 1 a kind of liver organization model configuration it is characterised in that: described protective layer (104) be close
Become macromolecular material.
4. a kind of liver organization model configuration as described in claim 1,2 or 3 it is characterised in that: described hydrogel is equal
The sodium alginate being 0.1～20% using mass percentage concentration, bioactive peptide, collagen, heparin, chondroitin sulfate, hyaluronic acid,
At least one in mucopolysaccharide, glycoprotein, matrigel, dextrose, shitosan, gelatin and fibrinogen solution.
5. as claimed in claim 4 a kind of liver organization model configuration it is characterised in that: described liver seed cell be liver thin
Born of the same parents, or stem cell regulating liver-QI cell mixture, or stem cell and growth factor mixture；Stem cell is fat stem cell, embryo does
At least one in cell and induced multi-potent stem cell；Described blood vessel seed cell be fat stem cell, mesenchymal stem cells MSCs,
At least one in vascular endothelial cell, smooth muscle cell, fibroblast, embryonic stem cell and induced multi-potent stem cell；Institute
Stating neural seed cell is neuron, or Schwann cell, or stem cell and growth factor mixture.
6. as claimed in claim 3 a kind of liver organization model configuration it is characterised in that: described synthesis macromolecular material is adopted
With in polyethylene, polycaprolactone, polyurethane, politef, polylactic acid, polyester and lactic acid and ethanol copolymer
Kind or the complex of different materials.
7. a kind of preparation method of liver organization model configuration as claimed in claim 1 it is characterised in that: the method include as
Lower step:
1) prepare particular manufacturing craft: the method using machining or 3 D-printing prepares described particular manufacturing craft, and this mould includes lower mould
(201), inner mold, simulation biliary system mould and mold；Described inner mold include simulate arterial vascular system inner mold,
Simulation venous vasculature inner mold and the inner mold of simulated nervous system；
2) by simulation tremulous pulse, vein and nervous system three inner molds (202) by the three of first order mold (203) top
Individual hole is inserted and fixed, and is then positioned on the first order step of lower mould (201) fixing together；Biliary system mould will be simulated
Tool (205) passes through the hole on lower mould (201) to fix；Afterwards the hydrogel containing blood vessel seed cell is filled into inner mold
(202) in the gap and first order mold (203) between, and the hydrogel containing blood vessel seed cell is made to hand over using cross-linking agent
Connection, removes first order mold (203), forms the stable hydrogel layer (107) containing blood vessel seed cell；
3) it is placed on the second level step of lower mould (201) solid using the diameter second level mold (204) bigger than the first order
Fixed, the hydrogel containing liver seed cell is filled into hydrogel layer (107) and the second level mold containing blood vessel seed cell
(204) in the gap between, and make the hydrogel containing liver seed cell crosslinked using cross-linking agent, remove second level mold
(204), form the stable hydrogel layer of seed cell containing liver (106)；
4) according to step 3) method, be placed on lower mould (201) using the diameter third level mold bigger than second level mold
Third level step on fixing, by the hydrogel containing stem cell be filled into liver seed cell hydrogel layer and third level mold it
Between gap in, and make hydrogel containing stem cell crosslinked using cross-linking agent, obtain the hydrogel layer containing stem cell；
5) remove inner mold and lower mould, irrigate respectively in the gap that simulation tremulous pulse and simulation venous vasculature inner mold stay
Vascular endothelial cell, forms arterial vascular system and simulation venous vasculature, the gap staying in simulated nervous system inner mold
Middle perfusion Schwann cell suspension, forms branch's nervous system；
6) exposed part biliary system outside agent structure is contained hepatocellular hydrogel parcel with one layer, and made using cross-linking agent
Celliferous hydrogel is crosslinked；
7) synthesis macromolecular material is dissolved in and in organic solvent, makes the synthesis macromolecular solution that mass percentage concentration is 5%～30%,
It is placed in spray bottle, become macromolecular material in agent structure surface even application one lamination shaping, be then placed in pbs molten
Extract in liquid, finally give stable liver organization model configuration.
8. a kind of preparation method of liver organization model configuration as claimed in claim 7 it is characterised in that: described mold,
Lower mould (201), simulation arterial vascular system inner mold (202a), simulation venous vasculature inner mold (202b), mould
Intend nervous system inner mold (202c) and simulation biliary system inner mold (205) adopts metal or synthesis macromolecular material；Described
Metal adopts aluminium alloy, pyrite or stainless steel material；Synthesis macromolecular material adopts politef, pbs plastics, photosensitive tree
Fat, rubber, acrylonitrile-butadiene-styrene copolymer plastics, polystyrene plastics, poly methyl methacrylate plastic, poly-
Formaldehyde plastics, polypropylene plastics, vinyon, polrvinyl chloride, Maranyl, polycarbonate plastic, epoxy resin,
Polycaprolactone, polyurethane or silicone plasties.
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| CN110403731A (en) * | 2019-07-30 | 2019-11-05 | 中国人民解放军陆军军医大学第一附属医院 | The bionical lobe of the liver structure of organizational project and preparation method based on living cells 3D printing |
| CN112111444A (en) * | 2020-08-28 | 2020-12-22 | 广东乾晖生物科技有限公司 | Method for reprogramming umbilical cord mesenchymal stem cells into liver cells and prepared liver organoid |
| CN113171490A (en) * | 2021-04-22 | 2021-07-27 | 中国医科大学 | Method for preparing hepatic precursor containing biliary tree structure by combining 3D printing and mold |
| CN113229993A (en) * | 2021-05-10 | 2021-08-10 | 中国医科大学 | Detachable combined die and method for preparing complex organ with multi-branch channel |
| CN113528338A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Special combined die for drug screening and use method thereof |
| CN113528337A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Combined die for organ manufacturing and drug screening and use method thereof |
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