CN102631710A - Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure - Google Patents
Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure Download PDFInfo
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- CN102631710A CN102631710A CN2012101103881A CN201210110388A CN102631710A CN 102631710 A CN102631710 A CN 102631710A CN 2012101103881 A CN2012101103881 A CN 2012101103881A CN 201210110388 A CN201210110388 A CN 201210110388A CN 102631710 A CN102631710 A CN 102631710A
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Abstract
The invention discloses a preparation method of a precursor of composite tissues and organs with a multichannel multilayer cell structure. The preparation method comprises the following steps that: one or more cell matrix solution and synthetic macromolecule solution are prepared, the synthetic macromolecule solution is injected into a main mold to form the lower surface of a support, and then the one or more cell matrix solution is injected into an assembling mold in a layered manner to form a plurality of cell matrix layers through crosslinking; and an inner mold is removed, the synthetic macromolecule solution is injected into the outer surface and the surrounding gap of the cell matrix layers to form an outer support, and the assembling mold is removed. The preparation method can be used for forming a complex multichannel three-dimensional structure combining different cell matrix materials containing uniform internal channels with a case of the synthetic macromolecule support, and overcomes the defects of tissue engineering that when cells are induced and cultured in the three-dimensional support, the needed time is long, the type of the cells is single, the distribution of the cells is nonuniform, the cells closed in a cell layer which exceeds a certain thickness are not easy to survive, nutrient channels do not exist, the layering differentiation on the stem cells is difficult to realize, the limitation on the appearance of the support is strong, and the like.
Description
Technical field
The invention belongs to the artificial manufacturing technology field of biological tissue and organ, particularly utilize synthesized polymer material, cell matrix materials to prepare the process of histoorgan precursor, belong to the bioengineered tissue technical field.
Background technology
Annual in the world patient's number of suffering from tissue defect or organ failure exceedes ten million.Yet the live body donor organ is limited, and the existing mechanical device does not possess all functions of organ, can not prevent that patient's the state of an illness from further worsening.In view of the above, arise at the historic moment with organizational project (Tissue Engineering) technology that to improve this type of illness treatment level be aim.
Tissue engineering is by the formal proposition and definite in 1987 of American National science fund committee; Be the principle of application cell biology, biomaterial and engineering, research and development are used for sick tissue or the structure of organ, the science of function of decreasing of gentrify human body.Organizational project is a new and high technology subject that is produced by multidisciplinary intersection such as biology, medical science, materialogy, engineering.Its implication is to use the philosophy and technique of life sciences and engineering; On the mammiferous normal and pathology two states organizational structure down of correct understanding and emic basis, study, develop the function that is used to repair, safeguard, promote behind various tissues of human body or the organ injury and the biological substitution thing of form.Wolter is formal " organizational project " speech that proposes in 1984, and American National science fund committee in 1987 confirms that formally tissue engineering becomes a new subject.The tissue engineering technique of scientists utilization for many years utilizes a small amount of normal cell of human body rudimentary organ to carry out external breeding, hopes to obtain patient's organ required, that have identical function, does not have rejection, has obtained gratifying achievement.
But existing tissue engineering technique faces many difficulties and restriction, and the success that the organizational project Applied Research Laboratory is obtained all is at the comparatively simple histoorgan of those structures and physiological function such as skeleton, cartilage, skin.The general preparation earlier of tradition Method of Tissue Engineering structure stand; In carrying out cell cultivation process because most oxygen of upper strata cell consumption and nutrition; Limited these components and spread, thereby limited cell to migration of support deep layer etc., the requirement that does not reach timely treatment clinical patient to bottom.Traditional single organization's support technology of preparing is difficult to form to be had the passage layered tissue of nutrition supply and can induce the differentiated tissues organ precursor.Traditional tissue engineering technique of while can not satisfy different cells accurately location and fixed point placement in the space, makes up the demand of the function gradient structure of complicated tissue organ.China has become organ transplantation second big country according to statistics; The patient who accepts organ transplantation every year is about 10,000 examples; But China has 1,500,000 routine needs of patients to carry out organ transplantation every year approximately, is badly in need of the new technology means and occurs in order to cultivate fast and effectively from the body organ in a large number.A kind of method for preparing of multichannel many cells complex tissue organ precursor can satisfy the crucial requirement in early stage of this great market just, and necessary preparation is provided for subsequently the complete organ growth of organizing.
Summary of the invention
The method for preparing that the purpose of this invention is to provide a kind of complex tissue organ precursor of multichannel multi-layer cellular structure; Be intended on the basis of previous work; Utilize assembling die to pour into step by step; Realize multi-layer cellular structure and timbering material in spatial accurate location, the complex tissue organ precursor of the multiple somatomedin of shaping multiple nutrient matter passage.Utilize principles such as die assembly, macromolecule solidification forming to realize the reconstruction of complicated tissue organ; The present invention can be shaped and contain the complex three-dimensional structure of different cell matrix materials and synthetic high polymer support; Overcome inducing culture cell in three-dimensional rack that organizational project exists need the time long, cell distribution is inhomogeneous, cell is difficult to penetrate into the medium shortcoming of endothecium structure.
Technical scheme of the present invention is following:
A kind of method for preparing of complex tissue organ precursor of multichannel multi-layer cellular structure is characterized in that this method comprises the steps to carry out:
1) synthesized polymer material is dissolved in the organic solvent, processes mass percentage concentration and be 20%～50% synthetic high polymer solution;
2) the preparation quality percentage concentration is 1%～30% multiple natural polymer solution respectively, with multiple natural polymer solution and various animals cell suspending liquid respectively by 1～9: 9～1 volume ratios are mixed and made into the various kinds of cell matrix solution; Cell concentration is 1 * 10 in the animal somatic cell suspension
4Individual/mL～1 * 10
7Individual/mL;
3) the pre-designed main mould and the inner mold of fitting with main mould circumferential inner surface; Said main mold bottom is provided with the club shaped structure of arranged in arrays; Inner mold is embedded main mould form assembling die; A part of synthetic high polymer NaOH solution tank NaOH is annotated in the assembling die, removed organic solvent, form bottom synthesized polymer material support through dry film method or wet film method;
4) one or more cellular matrix solution stratified are annotated in the assembling die,, made the natural polymer in the cellular matrix solution crosslinked, form the cellular matrix layer structure of multi-layer stable again through physics or Chemical Crosslinking Methods;
5) inner mold is removed from assembling die; Synthetic high polymer solution is poured into the slit that inner mold leaves over and the upper surface of multi-layer cellular hypothallus structure; Remove organic solvent through dry film method or wet film method; Remove main mould then, process the complicated tissue organ precursor of multichannel and multi-layer cellular structure.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure is characterized in that: described synthesized polymer material adopts the complex of one or more materials in polyurethane, lactic acid and ethanol copolymer, polylactic acid and the polyester.The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure is characterized in that: described natural macromolecular material adopts at least a in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and the FTN.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure; It is characterized in that: in cellular matrix solution, also add percent by volume and be 1%～30% frozen protective agent, described frozen protective agent adopts one or both mixtures of material in glycerol, dimethyl sulfoxide, ethylene glycol and the glucosan.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure is characterized in that: in cellular matrix solution, adding percent by volume is 0.001%～0.1% cell growth factor; Said somatomedin adopts one or more in ECGF, cell transfer factor and the hepatocyte growth factor.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure; It is characterized in that: said animal somatic cell adopts stem cell or becomes somatic cell; Said stem cell is fat stem cell, blood stem cell or bone marrow stem cell, and said one-tenth somatic cell is hepatocyte, myocardial cell or neurocyte.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure is characterized in that: be used to dissolve organic solvent employing TEG, ethylene glycol, the isopropyl alcohol or 1 of said synthesized polymer material in the step 1), the 4-dioxane; Step 2) solvent that is used to dissolve said natural macromolecular material in adopts 0.09M sodium chloride, 3-hydroxymethyl aminomethane hydrochloric acid solution or the cell culture fluid of water, normal saline, PBS solution, pH=6～8.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure; It is characterized in that: described main mould is processed by metal or hard macromolecular material; The cross sectional shape of described main mould and inner mold is circular, oval, polygon, or the structure of homologous organs surface configuration.
The synthetic high polymer timbering material possesses excellent mechanical performance in the complex tissue organ precursor that has multichannel multi-layer cellular structure that the present invention is prepared, and the structure that hole connects in the middle of the outside composite high-molecular material of this interior detail cytoplasmic matrix can be avoided simple cell substrate to implant being scattered and then by this difficult problem of autophagy.Wherein cellular matrix solution has excellent biocompatibility, and the multi-layer cellular structure can form multiple tissue therein.The present invention can realize multi-layer cellular structure/natural macromolecular material and synthetic high polymer timbering material in spatial accurate location, overcome inducing culture cell in three-dimensional rack that organizational project exists need the time long, cell category is single, cell distribution is inhomogeneous, cellular layer surpass certain thickness after airtight cell wherein be difficult for becoming works, no nutritional solution passage, stem cell layering to break up shortcomings such as difficult, that the contoured cradle limitation is strong.The present invention utilizes principles such as combination die general laws, macromolecule solidification forming to reach the requirement of different cell types of different parts in the complex organ and structure type, for the reconstruction that realizes complicated tissue organ lays the first stone.
Description of drawings
Fig. 1 is the schematic three dimensional views (is example with the cylindrical shape) of main mould and inner mold among the present invention.
Fig. 2 a is the front view (is example with the cylindrical shape) of assembling die among the present invention.
Fig. 2 b is the A-A cutaway view of Fig. 2 a.
Fig. 3 a is the front view (is example with the cylindrical shape) of main mould among the present invention.
Fig. 3 b is the A-A cutaway view of Fig. 3 a.
Fig. 4 is the section hierarchical diagram (is example with double-deck cellular matrix layer) of multichannel multi-layer cellular structure complex tissue organ precursor.
In Fig. 1 to Fig. 4:
The 1-inner mold; 2-master's mould; 3-bottom macromolecular material support; 4-cellular matrix layer structure.
The practical implementation method
The method for preparing of a kind of multichannel many cells complex tissue organ precursor provided by the invention, its concrete processing step is following:
1) synthesized polymer material is dissolved in the organic solvent, processes mass percentage concentration and be 20%～50% synthetic high polymer solution;
2) the preparation quality percentage concentration is 1%～30% multiple natural polymer solution respectively, with multiple natural polymer solution and various animals cell suspending liquid respectively by 1～9: 9～1 volume ratios are mixed and made into the various kinds of cell matrix solution; Cell concentration is 1 * 10 in the animal somatic cell suspension
4Individual/mL～1 * 10
7Individual/mL;
3) the pre-designed main mould 2 and the inner mold 1 of fitting with main mould circumferential inner surface; Said main mold bottom is furnished with the club shaped structure of array; The inner mold 1 embedded mould 2 of becoming owner of is formed assembling die; The synthetic high polymer NaOH solution tank NaOH is annotated in the assembling die, removed organic solvent, form bottom synthesized polymer material support 3 through dry film method or wet film method;
4) one or more cellular matrix solution stratified are annotated in the assembling die,, made the natural polymer in the cellular matrix solution crosslinked, form the cellular matrix layer structure 4 of multi-layer stable again through physics or Chemical Crosslinking Methods;
5) inner mold 1 is removed from assembling die; Again synthetic high polymer solution is poured into slit and the multi-layer cellular hypothallus structure upper surface that inner mold 1 is left over; Remove organic solvent through dry film method or wet film method; Remove main mould 2, process the complicated tissue organ precursor of multichannel and multi-layer cellular structure.
Preferred version of the present invention is that in described cellular matrix solution, also to add percent by volume be 1%～30% frozen protective agent, and described frozen protective agent adopts one or both mixtures of material in glycerol, dimethyl sulfoxide, ethylene glycol and the glucosan.In described cellular matrix solution, adding percent by volume is 0.001%～0.1% cell growth factor.Described cell growth factor adopts single or composite growth factors such as ECGF, cell transfer factor or hepatocyte growth factor.Described synthesized polymer material adopts the complex of one or more materials in polyurethane, lactic acid and ethanol copolymer, polylactic acid and the polyester.Described natural macromolecular material adopts at least a in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and the FTN.Said animal somatic cell adopts stem cell or becomes somatic cell, and said stem cell is fat stem cell, blood stem cell or bone marrow stem cell, and said one-tenth somatic cell is hepatocyte, myocardial cell or neurocyte.Be used to dissolve organic solvent employing TEG, ethylene glycol, the isopropyl alcohol or 1 of said synthesized polymer material in the step 1), the 4-dioxane; Step 2) solvent that is used to dissolve said natural macromolecular material in adopts 0.09M sodium chloride, 3-hydroxymethyl aminomethane hydrochloric acid solution or the cell culture fluid of water, normal saline, PBS solution, pH=6～8.
The method for preparing of the complex tissue organ precursor of described a kind of multichannel multi-layer cellular structure; It is characterized in that: described main mould 2 is processed by metal or hard macromolecular material; The cross sectional shape of described main mould 2 and inner mold 1 is circular, oval, polygon, or the structure of homologous organs surface configuration.
Embodiment 1:1) the pyrite material that do not wait of a series of sizes of the preparation mould that forms a complete set of; 2) PLGA/ one or four dioxane solution of preparation 50% (W/V), the heparin of adding 1% (W/W) pours in the main mould.Use the dry film method of natural air drying to form PLGA bottom support; 2) the preparation fibrinogen solution is become owner of mould inside with the inner mold cover, in assembling die, injects the mixture of gelatin/Fibrinogen and endotheliocyte, and cell density is 1 * 10
7Individual/mL; 3) add hepatocyte growth factor (HGF0.5ng/mL), human blood platelets derivation somatomedin (BB or PDGF-BB 50ng/mL), transforminggrowthfactor-(TGF β 1 10ng/mL) and basic fibroblast growth factor (b-FGF 2.5ng/mL).The cell natural macromolecular material is evenly distributed, injects thrombin solution (20IU/mL) and make cell/natural macromolecular material layer form rock-steady structure; 4) remove inner mold, leave over gap perfusion PLGA/ one or four dioxane solution, air-dry formation PLGA top layer and circumferential support at cell/natural macromolecular material layer upper surface and inner mold; 5) remove main mould, fix, accomplish the preparation of multichannel many cells complex tissue organ precursor with the stitching thread penetrating via.
Embodiment 2:1) prepares the mould that forms a complete set of of multichannel histoorgan precursor with silicone rubber; 2) polyurethane/ethylene glycol solution of preparation 25%, the paclitaxel of adding 5% stirs, and pours in the main mould.Use the cell culture liquid extraction method to form the outer polyurethane layer, be inserted in inner mold; 3) (cell density is 1 * 10 to prepare the Fibrinogen/endotheliocyte mixture that contains 5% paclitaxel
7Individual/as mL), to pour into to assembling die and add thrombin solution (20IU/mL), form the ground floor material of main part; 4) on the ground floor material of main part, inject the mixture of gelatin/Fibrinogen and fat stem cell, cell density is 1 * 10
6Individual/mL, form second layer material of main part; 5) (cell density is 1 * 10 on second layer material of main part, to inject Fibrinogen/hepatocyte mixture
7Individual/as mL), to soak the shaping thing 1 minute with thrombin solution (10IU/mL), form the three-decker material; 6) remove inner mold, leave over gap filling polyurethane/ethylene glycol solution at cell/natural macromolecular material layer upper surface and inner mold, extraction forms top layer and circumferential layer of polyurethane support through cell culture fluid; 7) remove main mould, coating can be induced and is the vessel growth factor at the passage place, and near tissue blood vesselization promoting is accomplished the preparation of implantable artificial liver precursor.
Embodiment 3:1) politef that do not wait of a series of sizes of the preparation mould that forms a complete set of; 2) compound concentration is polylactic acid/aqueous isopropanol of 30%, adds 30% sodium citrate, stirs, and pours in the main mould.Use the PBS extraction to form the bottom support, be inserted in inner mold; 3) (cell density is 1 * 10 to the collagen/endotheliocyte mixture of preparation 1% sodium citrate
7Individual/as mL), to pour into to assembling die, 37 ℃ of held 10 minutes make collagen/endotheliocyte mixture Stability Analysis of Structures, form the ground floor material of main part; 4) (cell density is 1 * 10 on the ground floor material of main part, to inject collagen/smooth muscle cell mixture
7Individual/mL); 37 ℃ of held 10 minutes make collagen/smooth muscle cell mixture Stability Analysis of Structures, form second layer material of main part; 5) mixture of injection collagen and fat stem cell/neonatal rat myocardial cell (1: 1) on second layer material of main part, cell density is 1 * 10
6Individual/mL, placed 10 minutes in 37 ℃ of incubators, form the 3rd layer main body material; 6) remove inner mold, leave over gap perfusion polylactic acid/aqueous isopropanol at cell/natural macromolecular material layer upper surface and inner mold, extraction forms top layer and circumferential support through PBS; 7) remove main mould, accomplish the preparation of implantable artificial heart precursor.
Embodiment 4:1) prepares supporting main mould and inner mold with politef; 2) preparation 30%PU/ TEG solution pours in the main mould.Use the PBS extraction to form the bottom support, be inserted in inner mold; 3) prepare following solution: two kinds of natural biologic materials of Fibrinogen and gelatin are dissolved in respectively in phosphate buffer (PBS) solution processes 10% and 30% macromolecular solution, even by 1: 1 (v/v) mixed again.Add 10% dimethyl sulfoxide, 5% glucosan then by volume; Fat stem cell and messangial cell is even by 1: 1 mixed, add in the macromolecular solution, (cell density is 1 * 10 to obtain fat stem cell-messangial cell-gelatin-Fibrinogen-dimethyl sulfoxide-glucosan mixture
4Individual/as mL), to pour into to assembling die, and with thrombin solution (30IU/mL) fixing 2 minutes; 4) remove inner mold, leave over gap perfusion PU/ TEG solution,, form top layer and circumferential support through the PBS extractant at cell/natural macromolecular material layer upper surface and inner mold; 5) remove main mould, above-mentioned three-dimensional structure 4 ℃ of held half an hour, is placed on half an hour in-20 ℃ of refrigerators, put into-70 ℃ of cryogenic refrigerator liquid nitrogen cryopreservation at last, rapid rewarming during use is cultivated in order to using.
Embodiment 5:1) the pyrite material that do not wait of a series of sizes of the preparation mould that forms a complete set of; 2) preparation contains the 30% polyester/TEG solution of 3% paclitaxel, pours in the main mould, through the PBS extractant, forms the bottom support, is inserted in inner mold; 3) Fibrinogen is dissolved in phosphate buffer (PBS) solution processes 10% macromolecular solution.Add 20% glycerol, 5% glucosan then by volume; Fat stem cell and islet cells is even by 2: 1 mixed, and (cell density is 1 * 10 in the adding macromolecule mixed solution
7Individual/as mL), to obtain fat stem cell-islet cells, gelatin-Fibrinogen-dimethyl sulfoxide-glucosan mixture, pour into to assembling die, fix 2 minutes with thrombin solution (10IU/mL); 4) remove inner mold, leave over gap perfusion polyester/TEG solution,, form top layer and circumferential support through the PBS extractant at cell/natural macromolecular material layer upper surface and inner mold; 5) remove main mould, above-mentioned three-dimensional structure 4 ℃ of held half an hour, is placed on half an hour in-20 ℃ of refrigerators, put into-196 ℃ of liquid nitrogen cryopreservation at last, rapid rewarming during use adds culture fluid in 37 ℃, 5%CO
2Cultivate subsequent use under the condition.
Claims (8)
1. the method for preparing of the complex tissue organ precursor of a multichannel multi-layer cellular structure is characterized in that this method comprises the steps to carry out:
1) synthesized polymer material is dissolved in the organic solvent, processes mass percentage concentration and be 20%～50% synthetic high polymer solution;
2) the preparation quality percentage concentration is 1%～30% multiple natural polymer solution respectively, with multiple natural polymer solution and various animals cell suspending liquid respectively by 1～9: 9～1 volume ratios are mixed and made into the various kinds of cell matrix solution; Cell concentration is 1 * 10 in the animal somatic cell suspension
4Individual/mL～1 * 10
7Individual/mL;
3) the pre-designed main mould (2) and the inner mold (1) of fitting with main mould circumferential inner surface; Said main mold bottom is provided with the club shaped structure of arranged in arrays; Inner mold (1) is embedded main mould (2) form assembling die; A part of synthetic high polymer NaOH solution tank NaOH is annotated in the assembling die, removed organic solvent, form bottom synthesized polymer material support (3) through dry film method or wet film method;
4) one or more cellular matrix solution stratified are annotated in the assembling die,, made the natural polymer in the cellular matrix solution crosslinked, form the cellular matrix layer structure (4) of multi-layer stable again through physics or Chemical Crosslinking Methods;
5) inner mold (1) is removed from assembling die; Synthetic high polymer solution is poured into the slit that inner mold (1) leaves over and the upper surface of multi-layer cellular hypothallus structure; Remove organic solvent through dry film method or wet film method; Remove main mould (2) then, process the complicated tissue organ precursor of multichannel and multi-layer cellular structure.
2. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 1, it is characterized in that: described synthesized polymer material adopts the complex of one or more materials in polyurethane, lactic acid and ethanol copolymer, polylactic acid and the polyester.
3. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 1, it is characterized in that: described natural macromolecular material adopts at least a in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and the FTN.
4. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 1; It is characterized in that: in cellular matrix solution, also add percent by volume and be 1%～30% frozen protective agent, described frozen protective agent adopts one or both mixtures of material in glycerol, dimethyl sulfoxide, ethylene glycol and the glucosan.
5. according to the method for preparing of the complex tissue organ precursor of claim 1 or 4 described a kind of multichannel multi-layer cellular structures, it is characterized in that: in cellular matrix solution, adding percent by volume is 0.001%～0.1% cell growth factor; Said somatomedin adopts one or more in ECGF, cell transfer factor, human blood platelets derivation somatomedin, transforming growth factor, basic fibroblast growth factor and the hepatocyte growth factor.
6. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 5; It is characterized in that: said animal somatic cell adopts stem cell or becomes somatic cell; Said stem cell is fat stem cell, blood stem cell or bone marrow stem cell, and said one-tenth somatic cell is hepatocyte, myocardial cell or neurocyte.
7. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 1; It is characterized in that: be used to dissolve organic solvent employing TEG, ethylene glycol, the isopropyl alcohol or 1 of said synthesized polymer material in the step 1), the 4-dioxane; Step 2) solvent that is used to dissolve said natural macromolecular material in adopts 0.09M sodium chloride, 3-hydroxymethyl aminomethane hydrochloric acid solution or the cell culture fluid of water, normal saline, PBS solution, pH=6～8.
8. according to the method for preparing of the complex tissue organ precursor of the described a kind of multichannel multi-layer cellular structure of claim 1; It is characterized in that: described main mould (2) is processed by metal or hard macromolecular material; The cross sectional shape of described main mould (2) and inner mold (1) is circular, oval, polygon, or the structure of homologous organs surface configuration.
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| CN103710263A (en) * | 2012-09-28 | 2014-04-09 | 江苏吉锐生物技术有限公司 | Cell culture apparatus |
| CN105343937A (en) * | 2015-11-10 | 2016-02-24 | 武汉华一同信生物科技有限公司 | Composite porous sponge material and preparation method thereof |
| CN107929808A (en) * | 2017-10-11 | 2018-04-20 | 西南大学 | A kind of preparation method of calcium alginate nerve trachea and Nerve Scaffold |
| CN109486675A (en) * | 2018-10-19 | 2019-03-19 | 天津大学 | A kind of skull vascular compound rest dynamic device for casting |
| CN111139213A (en) * | 2020-01-06 | 2020-05-12 | 清华大学 | Multilayer structure stent and preparation method and application thereof |
| CN112154201A (en) * | 2018-05-22 | 2020-12-29 | 形态细胞科技公司 | Perfusion bioreactors, perfusion devices, artificial liver systems, and related methods |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103710263A (en) * | 2012-09-28 | 2014-04-09 | 江苏吉锐生物技术有限公司 | Cell culture apparatus |
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| CN105343937A (en) * | 2015-11-10 | 2016-02-24 | 武汉华一同信生物科技有限公司 | Composite porous sponge material and preparation method thereof |
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| CN107929808B (en) * | 2017-10-11 | 2020-09-29 | 西南大学 | Preparation method of calcium alginate nerve conduit and nerve scaffold |
| CN112154201A (en) * | 2018-05-22 | 2020-12-29 | 形态细胞科技公司 | Perfusion bioreactors, perfusion devices, artificial liver systems, and related methods |
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| CN111139213B (en) * | 2020-01-06 | 2022-02-01 | 清华大学 | Multilayer structure stent and preparation method and application thereof |
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Application publication date: 20120815 |