CN107058216A - A kind of method for promoting articular cartilage tissue to grow - Google Patents

A kind of method for promoting articular cartilage tissue to grow Download PDF

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CN107058216A
CN107058216A CN201610887427.7A CN201610887427A CN107058216A CN 107058216 A CN107058216 A CN 107058216A CN 201610887427 A CN201610887427 A CN 201610887427A CN 107058216 A CN107058216 A CN 107058216A
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articular cartilage
tissue
culture
cartilage tissue
biometric print
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崔晓峰
高桂芳
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Wuhan Feng Lin Technology Co Ltd
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Wuhan Feng Lin Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0655—Chondrocytes; Cartilage
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y10/00—Processes of additive manufacturing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/113—Acidic fibroblast growth factor (aFGF, FGF-1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/119—Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/148—Transforming growth factor alpha [TGF-a]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/15—Transforming growth factor beta (TGF-β)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2513/00—3D culture

Abstract

The invention discloses a kind of method for promoting articular cartilage tissue to grow, comprise the following steps：a）Human articular cartilage sample is taken, cartilage fragment is digested with trypsase familial combined hyperlipidemia Clostridial collagenase respectively, washed immediately after digestion；b）The human articular chondrocytes discharged are washed with DMEM, inoculated and cultured is carried out to the articular chondrocytes of separation；c）Bio-ink is prepared using the articular chondrocytes of culture, polyethylene glycol dimethylacrylate and PBS；d）Printing builds 3D hydrogel soft bone tissues on biometric print paper；e）3D hydrogel soft bone tissues are cultivated in the medium, form articular cartilage tissue.What the present invention was provided promotes the method technique of articular cartilage tissue growth advanced, easy to operate, repeatable strong, the usage amount of human chondrocytes can effectively be reduced, usage amount is reduced up to 50%, while effectively facilitating the growth of function cartilaginous tissue, accelerates tissue growth to be up to 40%.

Description

A kind of method for promoting articular cartilage tissue to grow
Technical field
The invention belongs to biology and organizational engineering technical field, more particularly to a kind of promotion articular cartilage tissue growth Method.
Background technology
Cartilaginous tissue mainly plays support, protection, dispersive stress etc., can divided in histology as a kind of connective tissue For hyaline cartilage, elastic cartilage, the class of fibrocartilage three.They are all histologically the single organizations for comprising only cartilage cell, Inside is without blood vessel, and nutrition relies primarily on the infiltration of tissue fluid.Articular cartilage damage caused by the disease such as wound and Osteoarthritis One of subject matter for being posed a health risk in global range is still with degenerative disease.Although operation or No operation measure intervention are controlled Made much progress in terms for the treatment of, but the reparation of cartilage damage is still very stubborn problem.In recent years, the appearance of organizational project is soft The reparation of bone injury provides a kind of new, effective treatment means.
Biometric print based on thermal inkjet-printing technology was suggested in 2003 and for biological manufacture, with high flux, It is digital control, place cell, biotic factor and the advantages of biologic bracket material in two-dimensional/three-dimensional energy pin-point accuracy.This technology is There are many successfully precedents.By biological 3D printing and to plant the cartilage cell of people come external structure function articular cartilage be a kind of Great scientific research and the method for clinical treatment meaning.Clinically a significant challenge of autogenous cell repair of cartilage is exactly limited Live body cartilage cell's resource, in the urgent need to one kind can after biometric print effective amplifying cells, while can guarantee that outer base again The method of matter quality.It is of greatest concern at present be exactly Autologous Chondrocyte or mescenchymal stem cell that directly transplanting is enriched with without With amplification in vitro, and in order to play to greatest extent, biometric print is accurate in 3D structures to place cell, growth factor, biology The advantage of timbering material, limited initial cell density will optimize the placement degree of accuracy.
In summary, cartilage tissue engineered field is in the urgent need to developing a kind of method of new constructing function cartilage, a side Face can effectively reduce the usage amount of cartilage cell, on the other hand can effectively facilitate the growth of cartilaginous tissue.
The content of the invention
In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to which cell consumption and energy can be greatly reduced by now providing one kind Substantially speed up the method for promoting articular cartilage tissue growth of tissue growth.
In order to solve the above technical problems, the technical solution adopted by the present invention is：It is a kind of to promote what articular cartilage tissue grew Method, its innovative point is：Comprise the following steps：a）Human articular cartilage sample is taken, cartilage fragment is cut into, tryptose is used respectively Enzyme familial combined hyperlipidemia Clostridial collagenase digests to cartilage fragment, is washed immediately after digestion；b）With containing 1% (v/v) 1 × penicillin-chain The articular chondrocytes of separation are carried out inoculation training by the human articular chondrocytes that the DMEM washings of mycin-glutamine are discharged Support；c）Bio-ink is prepared using the articular chondrocytes of culture, polyethylene glycol dimethylacrylate and PBS；d）Make Structure 3D hydrogel soft bone tissues are printed on biometric print paper with biometric print machine；e）3D hydrogel softs are cultivated in the medium Bone tissue, forms articular cartilage tissue.
Further, the step a）Middle digestion process specific steps include：Cartilage fragment is first placed in 0.5mg/ml's In trypsase, 15min is digested at 37 DEG C, then, trypsase is removed, then be placed in the IV type clostridium collagen containing 2mg/ml Enzyme, 5%（v/v）In the DMEM of hyclone, 12-16h is digested at 37 DEG C.
Further, the step b）Middle washing times are 3 times；Cell viability after washing is more than 95%；The joint of separation Cartilage cell is inoculated in T175 tissue culture flasks, carries out individual layer amplification cultivation, and inoculum concentration is every bottle of 5,000,000 cells, culture Base is the DMEM containing 10% (v/v) calf serum, 1% (v/v) 1 × Pen .- Strep-glutamine, and cultivation temperature is 37 DEG C, Culture environment is 5% (v/v) CO2 wet air, and culture medium is changed once for every 4 days.
Further, the step c）The middle specific steps for preparing bio-ink include：Take a certain amount of polyethylene second two Alcohol dimethylacrylate is dissolved in the solution that final concentration of 10% (w/v) is formed in PBS, and adding light trigger 2959 makes solution Concentration is 0.5% (w/v), and cultured articular chondrocytes are suspended in solution, and cell density is 8 × 106cells/ml。
Further, the step d）Middle biometric print machine prints structure 3D hydrogel soft bone tissues on biometric print paper Specific steps include：Preparation before the printing of biometric print machine；The bio-ink prepared is loaded into printing pen and tinfoil paper is used Paper bag is lived；Use print software design object model and moulded dimension；Printing builds 3D hydrogels layer by layer on biometric print paper Cartilaginous tissue.
Further, preparation includes before the biometric print machine printing：Biometric print machine is degerming, by biometric print machine It is degerming with more than 2h is irradiated under ultraviolet；Pen cleaning is printed, is first sterilized with 70% (v/v) alcohol flushing, then use aseptic water washing Totally；Long wave ultraviolet lamp is installed, long wave ultraviolet lamp being placed in above print platform at 25cm, light is completed during for printing Polymerisation.
Further, the step d）Middle biometric print paper is set to diameter 4mm cylindrical die, itself and printhead Distance is 1-2mm；During printing, the intensity for keeping ultraviolet on print platform is 4-8mW/cm2。
Further, the step e）The incubation time of middle 3D hydrogel softs bone tissue in the medium is 4 weeks；Culture temperature Degree is 37 DEG C, and culture environment is 5% (v/v) CO2 wet air, and culture medium is changed once for every 3 days；Culture first week, is used The ITS+ culture mediums of fibroblast growth factor/TGF supplement are cultivated, and are cultivated the 2-4 weeks, are used conversion The ITS+ culture mediums of growth factor supplement are cultivated.
Further, into fiber in the ITS+ culture mediums of the fibroblast growth factor/TGF supplement The content of Porcine HGF/TGF is 10ng/ml, in the ITS+ culture mediums of the TGF supplement The content of TGF is 10ng/ml.
Beneficial effects of the present invention are as follows：
What 1. the present invention was provided promotes the method technique of articular cartilage tissue growth advanced, easy to operate, repeatable strong, can The effectively usage amount of reduction human chondrocytes, usage amount reduction reachable 50%, while the growth of function cartilaginous tissue is effectively facilitated, Accelerate tissue growth to be up to 40%, can be provided for the clinical treatment of cartilage tissue engineered scientific research and cartilage damage Beneficial reference, effective technical support is provided for the clinicization of biological printing technique.
2. loading the bio-ink prepared after printhead in the present invention, encased with masking foil, be effectively prevented from biology Cell in ink is by ultraviolet injury.
Brief description of the drawings
Joint is formed by 3D hydrogel soft bone tissue cultures in the method that Fig. 1 produces for a kind of promotion cartilaginous tissue of the present invention The expression conditions of first, second, third and fourth week interior NTx during cartilaginous tissue.
Joint is formed by 3D hydrogel soft bone tissue cultures in the method that Fig. 2 produces for a kind of promotion cartilaginous tissue of the present invention The expression conditions of first, second, third and fourth week interior proteoglycans during cartilaginous tissue.
Joint is formed by 3D hydrogel soft bone tissue cultures in the method that Fig. 3 produces for a kind of promotion cartilaginous tissue of the present invention The expression conditions of first, second, third and fourth week interior II Collagen Type VI during cartilaginous tissue.
Embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation Content disclosed by book understands other advantages and effect of the present invention easily.
A kind of method for promoting articular cartilage tissue to grow, comprises the following steps：
（1）The separation and culture of human chondrocytes
Articular cartilage sample is separated from corpse of the death time in 72 hours and obtained.Collection obtain cartilage samples shred it is rearmounted Digest 15 minutes, be placed in it again containing the type shuttles of 2mg/ml IV after removing trypsase at 37 DEG C in 0.5mg/ml trypsase Bacterium clostridiopetidase A, 5%（v/v）In the DMEM of hyclone, 12-16h is digested at 37 DEG C.
With 1%（v/v）The human chondrocytes three that the DMEM washings of 1 × Pen .- Strep-glutamine supplement are discharged Secondary, cell viability is more than 95% after washing.The cartilage cell of separation is inoculated in T175 tissue culture flasks and carries out individual layer amplification training Support, inoculum concentration is every bottle of 5,000,000 cells.Culture medium is containing 10% (v/v) calf serum, 1% (v/v) 1 × penicillin-strepto- The DMEM of element-glutamine.Condition of culture is 37 DEG C, 5% (v/v) CO2 wet air, and culture medium is changed once for every four days.When Cell reaches can be used to prepare bio-ink during 80-90% degrees of fusion.
（2）Prepare bio-ink
Appropriate polyethylene glycol dimethylacrylate is taken to be dissolved in PBS, final concentration of 10 % (w/v) of solution adds light Initiator 2959 makes solution concentration be 0.5 % (w/v), and cultured human articular chondrocytes are suspended in into 0.5 % (w/v) solution In, cell density is 8 × 10 in solution6cells/ml。
（3）Biometric print external structure 3D hydrogel soft bone tissues
Before printing, biometric print machine is placed under ultraviolet irradiate at least 2 hours it is degerming；Pen will be printed first with 70% (v/v) ethanol Flush three times again clean with aseptic water washing；A long wave ultraviolet lamp is placed above print platform at 25cm, in printing While complete photopolymerization reaction；It is 4-8mW/cm2 that the uitraviolet intensity on platform is kept during printing.By the biology ink prepared Water loads printing pen and encased with masking foil, it is to avoid cell is by ultraviolet injury.Biometric print paper is diameter 4mm cylinder Mould, the distance of itself and printhead is 1-2mm.Using Adobe Photoshop Software for Design object modules and its size, pass through Printing builds 3D hydrogel soft bone tissues layer by layer.
（4）The culture of articular cartilage tissue
Printed 3D hydrogel soft bone tissues are placed in culture medium and cultivated, temperature is 37 DEG C, and culture environment is 5%（v/v） CO2 wet air, culture medium is changed once for every 3 days, forms cartilaginous tissue.Wherein, cultivate first week use 10ng/ml into The ITS+ culture mediums of fibroblast growth factor/TGF supplement, must be converted for subsequent the 2-4 weeks using 10ng/ml The ITS+ culture mediums of growth factor supplement, the culture medium is standard Subchondral drilling culture medium.Fibroblast growth factor/turn Change growth factor supplement ITS+ culture mediums be in ITS+ culture mediums increase Basic fibroblast growth factor and conversion growth because Sub- mixture is used as supplement.
What the present invention was provided promotes the method technique of articular cartilage tissue growth advanced, easy to operate, repeatable strong, energy The usage amount of enough effectively reduction human chondrocytes, usage amount is reduced up to 50%, while effectively facilitating the life of function cartilaginous tissue It is long, accelerate tissue growth to be up to 40%, can be provided for the clinical treatment of cartilage tissue engineered scientific research and cartilage damage Beneficial reference, for the clinicization of biological printing technique provides effective technical support.
In order to further appreciate that the effect of the inventive method, corresponding identification has been carried out to turning out the cartilaginous tissue come.
（1）Real-time PCR Analysis cartilage cell's gene expression
The gene expression of cell in first, second, third and fourth week analysis hydrogel of articular cartilage tissue incubation.Analysis side Method for will turn out the cartilaginous tissue come be put into freezed in liquid nitrogen after crush, will it is broken after powder be transferred in centrifuge tube, to Appropriate RNA lysis buffers are added in centrifuge tube, are digested 15 minutes at room temperature；Then the solution of formation is transferred to another centrifugation Guan Zhong, is centrifuged 2 minutes with 12000g rotating speed, and RNA is extracted in supernatant and is purified.Use model Nanodrop ND- 1000 ultraviolet specrophotometer determines total RNA content and purity.Using the cDNA reverse transcription reagent box of high power capacity by separation Total serum IgE obtains cDNA through reverse transcription, and RT-PCR detection people NTx, II type are utilized using TaqMan gene expression detections probe The gene expression of collagen and proteoglycans, as shown in Figure 1, Figure 2 and Figure 3.
（2）Biochemistry detection
It is to take the cartilaginous tissue turned out 1,2,3,4 weeks of culture respectively in different timing nodes, and it is small to be lyophilized 48 When, afterwards with scalpel will it is lyophilized after cartilaginous tissue thinly slice and be respectively placed in papain and pepsin in digest.Often Individual sample extracts 16h for the pawpaw enzyme solutions of 125 mcg/mls using 1 milliliter of concentration at 60 DEG C and obtains total glycosaminoglycan, often Individual sample uses 1 milliliter of pepsin solution（Containing 100 mcg/ml pepsins）II Collagen Type VI is obtained in digestion at 4 DEG C within 6 days Enzyme.Human cartilage of the total glycosaminoglycan, II Collagen Type VI content and DNA content, the ratio of dry weight of each sample to assess implantation The biosynthesis activity of cell, the DNA content of each sample and the ratio of dry weight are to assess the propagation of the cell in incubation.
(3) histology biopsy
According to Standard histological detection method, the cartilaginous tissue for turning out is placed in 10% (v/v) formalin and fixed At night, it is then taken out being transferred in PBS up to being embedded into paraffin, paraffin section is carried out afterwards, slice thickness is 6 microns. Finally dyed with safranin O/fast green section to above-mentioned preparation, cell and proteoglycans in observation sample.
Above-described embodiment is presently preferred embodiments of the present invention, is not the limitation to technical solution of the present invention, as long as The technical scheme that can be realized without creative work on the basis of above-described embodiment, is regarded as falling into patent of the present invention Rights protection scope in.

Claims (9)

1. a kind of method for promoting articular cartilage tissue to grow, it is characterised in that：Comprise the following steps：a）Take human articular cartilage Sample, is cut into cartilage fragment, cartilage fragment is digested with trypsase familial combined hyperlipidemia Clostridial collagenase respectively, after digestion immediately Washing；b）The human articular chondrocytes discharged are washed with the DMEM containing 1% (v/v) 1 × Pen .- Strep-glutamine, Inoculated and cultured is carried out to the articular chondrocytes of separation；c）Using the articular chondrocytes of culture, polyethylene glycol dimethyl Acrylate and PBS prepare bio-ink；d）Structure 3D hydrogel cartilages are printed on biometric print paper using biometric print machine Tissue；e）3D hydrogel soft bone tissues are cultivated in the medium, form articular cartilage tissue.
2. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step a） Middle digestion process specific steps include：First cartilage fragment is placed in 0.5mg/ml trypsase, digested at 37 DEG C 15min, then, removes trypsase, then be placed in the IV type Clostridial collagenase containing 2mg/ml, 5%（v/v）The DMEM of hyclone In, digest 12-16h at 37 DEG C.
3. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step b） Middle washing times are 3 times；Cell viability after washing is more than 95%；The articular chondrocytes of separation are inoculated in T175 tissue cultures In bottle, individual layer amplification cultivation is carried out, inoculum concentration is every bottle of 5,000,000 cells, and culture medium is containing 10% (v/v) calf serum, 1% (v/v) DMEM of 1 × Pen .- Strep-glutamine, cultivation temperature is 37 DEG C, and culture environment is wet for 5% (v/v) CO2's Moisten air, culture medium is changed once for every 4 days.
4. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step c） The middle specific steps for preparing bio-ink include：A certain amount of polyethylene glycol dimethylacrylate is taken to be dissolved in shape in PBS Into final concentration of 10% (w/v) solution, adding light trigger 2959 makes solution concentration be 0.5% (w/v), is closed cultured Section cartilage cell is suspended in solution, and cell density is 8 × 106cells/ml。
5. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step d） The specific steps that middle biometric print machine prints structure 3D hydrogel soft bone tissues on biometric print paper include：Biometric print machine is beaten Preparation before print；The bio-ink prepared is loaded in printing pen and encased with masking foil；Mesh is designed using print software Mark model and moulded dimension；Printing builds 3D hydrogel soft bone tissues layer by layer on biometric print paper.
6. a kind of method for promoting articular cartilage tissue to grow according to claim 5, it is characterised in that：The biology is beaten Preparation includes before the printing of print machine：Biometric print machine is degerming, degerming by more than 2h is irradiated under biometric print machine ultraviolet； Pen cleaning is printed, is first sterilized with 70% (v/v) alcohol flushing, then it is clean with aseptic water washing；Long wave ultraviolet lamp is installed, by length Ripple ultraviolet lamp is placed in above print platform at 25cm, and photopolymerization reaction is completed during for printing.
7. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step d） Middle biometric print paper is set to diameter 4mm cylindrical die, and the distance of itself and printhead is 1-2mm；During printing, printing is kept The intensity of ultraviolet is 4-8mW/cm on platform2。
8. a kind of method for promoting articular cartilage tissue to grow according to claim 1, it is characterised in that：The step e） The incubation time of middle 3D hydrogel softs bone tissue in the medium is 3 weeks；Cultivation temperature is 37 DEG C, and culture environment is 5% (v/ V) CO2 wet air, culture medium is changed once for every 3 days；Culture first week, is given birth to using fibroblast growth factor/conversion The ITS+ culture mediums of long factor supplement are cultivated, and are cultivated the 2-3 weeks, the ITS+ culture mediums supplemented using TGF are entered Row culture.
9. a kind of method for promoting articular cartilage tissue to grow according to claim 8, it is characterised in that：It is described into fiber Fibroblast growth factor/TGF in the ITS+ culture mediums of Porcine HGF/TGF supplement Content be that the content of TGF in 10ng/ml, the ITS+ culture mediums of TGF supplement is 10ng/ ml。
CN201610887427.7A 2016-10-12 2016-10-12 A kind of method for promoting articular cartilage tissue to grow Pending CN107058216A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109822889A (en) * 2019-03-22 2019-05-31 河北雄安大洲智造科技有限公司 A kind of multichannel 3D printing spray head and preparation method thereof, cell/bio-ink transportation system
CN112386743A (en) * 2020-11-18 2021-02-23 华中科技大学同济医学院附属同济医院 Artificial ovary stent constructed by 3D biological printing technology, artificial ovary and construction method and application thereof
CN114699559A (en) * 2022-03-18 2022-07-05 北京大学第三医院（北京大学第三临床医学院） Construction method of cartilage scaffold

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CUI XF: "Direct Human Cartilage Repair Using Three-Dimensional Bioprinting Technology", 《TISSUE ENGINEERING PART A》 *
CUI X等: "Synergistic Action of Fibroblast Growth Factor-2 and Transforming Growth Factor-Beta1 Enhances Bioprinted Human Neocartilage Formation", 《BIOTECHNOL BIOENG》 *
GAO GF等: "Improved properties of bone and cartilage tissue from 3D inkjet-bioprinted human mesenchymal stem cells by simultaneous deposition and photocrosslinking in PEG-GelMA", 《BIOTECHNOLOGY LETTERS》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109822889A (en) * 2019-03-22 2019-05-31 河北雄安大洲智造科技有限公司 A kind of multichannel 3D printing spray head and preparation method thereof, cell/bio-ink transportation system
CN109822889B (en) * 2019-03-22 2024-03-26 深圳大洲医学科技有限公司 Multichannel 3D printing spray head, preparation method thereof and cell/biological ink conveying system
CN112386743A (en) * 2020-11-18 2021-02-23 华中科技大学同济医学院附属同济医院 Artificial ovary stent constructed by 3D biological printing technology, artificial ovary and construction method and application thereof
CN114699559A (en) * 2022-03-18 2022-07-05 北京大学第三医院（北京大学第三临床医学院） Construction method of cartilage scaffold

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Application publication date: 20170818