CN113061566A - Cell large-scale culture method and microcarrier used by same - Google Patents
Cell large-scale culture method and microcarrier used by same Download PDFInfo
- Publication number
- CN113061566A CN113061566A CN202110321555.6A CN202110321555A CN113061566A CN 113061566 A CN113061566 A CN 113061566A CN 202110321555 A CN202110321555 A CN 202110321555A CN 113061566 A CN113061566 A CN 113061566A
- Authority
- CN
- China
- Prior art keywords
- microcarrier
- cells
- solution
- hydrogel
- molecule
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
- C12N5/0068—General culture methods using substrates
- C12N5/0075—General culture methods using substrates using microcarriers
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
- C12N2500/10—Metals; Metal chelators
- C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
-
- 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
- C12N2531/00—Microcarriers
-
- 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
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/70—Polysaccharides
- C12N2533/72—Chitin, chitosan
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The application discloses a microcarrier, which comprises gel molecules and an outer matrix molecule, wherein the gel molecules and the outer matrix molecule are mixed and then induced to carry out a gelation reaction to generate a hydrogel microcarrier; wherein the gel molecule is sodium alginate or calcium alginate, and the outer matrix molecule is one or more of collagen, matrigel protein, chondroitin sulfate, fibronectin and chitosan. The main component of the microcarrier in the application contains calcium alginate composite. The material has the greatest advantage that the material is easy to expand and deform in the presence of the calcium ion chelating agent, so that the contact area between the material and cells is reduced, the cells can fall off under slight physical shearing force, and the separation of the cells and microcarriers is realized. The whole process can realize cell separation without the participation of pancreatin.
Description
Technical Field
The application relates to the technical field of cell culture, in particular to a cell large-scale culture method and a used microcarrier.
Background
With the development of stem cell technology, cell therapy offers the possibility of radical treatment of many major diseases such as tumors, liver and kidney organ failure, neuro-endocrine system diseases such as parkinson, diabetes, and the like. However, the cell product used for clinical implantation needs to avoid the introduction of various proteins and enzymes in the preparation process, which brings difficulties to the large-scale cell culture work. Especially aiming at the large-scale culture of the anchorage dependent cells, the pancreatin digestion is needed to collect the cultured cells, the introduction of the pancreatin increases the subsequent process flow of cell product purification, and increases the production cost of the product.
Disclosure of Invention
The purpose of the application is to overcome the defects of the prior art and provide a cell large-scale culture method without trypsinization and a microcarrier for realizing the culture method.
In order to achieve the technical purpose, the technical scheme adopted by the application is as follows:
in a first aspect, a microcarrier comprises a gel molecule and an extracellular matrix molecule, wherein the gel molecule and the extracellular matrix molecule are mixed and then induce a gelation reaction to produce a hydrogel microcarrier; wherein the gel molecule is sodium alginate or calcium alginate, and the outer matrix molecule is one or more of collagen, matrigel protein, chondroitin sulfate, fibronectin and chitosan.
The dry powder of the hydrogel microcarrier can swell and deform when soaked in a solution containing a sodium ion and calcium ion chelating agent.
Specifically, the calcium ion chelating agent is any one of phosphate, carbonate, bicarbonate, citrate or EDTA.
In a second aspect, there is provided a method for preparing a microcarrier, which comprises the steps of:
respectively preparing a gel molecule solution and an outer matrix molecule solution;
mixing calcium carbonate powder with the gel molecular solution, and then mixing with the outer matrix molecular solution to form a premixed solution;
promoting the pre-mixed liquid to carry out gelation reaction by a fluid granulation technology to generate a hydrogel microcarrier;
and drying and sterilizing the hydrogel microcarrier to obtain dry powder of the microcarrier.
Preferably, the concentration of the gel molecule solution is 10-50g/L, and the concentration of the outer matrix molecule solution is 1-50 g/L.
More preferably, the concentration of the gel molecule solution in the premix is 10g/L or more.
Specifically, the fluid granulation technology is any one of a high-voltage electrostatic field technology, a gas orifice spraying technology, a micro-fluidic technology or an emulsification technology.
Further, after the hydrogel microcarrier is generated, adding 1-10g/L of chitosan solution, adding the outer matrix molecule solution, and performing polyelectrolyte complexation reaction to generate the hydrogel microcarrier with a shell-core structure. The volume ratio of the hydrogel microcarrier to the chitosan solution is 1: 2-10; the duration time of the polyelectrolyte complexation reaction is 2-60 minutes.
Further, after the hydrogel microcarrier is formed, the hydrogel microcarrier is placed in an acidic environment to produce carbon dioxide, resulting in a hydrogel microcarrier having a porous structure.
In a third aspect, a method for large-scale cell culture comprises the following steps:
preparing microcarrier and seed cell separately;
co-culturing the microcarrier and the seed cells by using a serum-free culture medium which does not contain a sodium ion and calcium ion chelating agent, so that the seed cells are adhered to the surface of the microcarrier for proliferation;
replacing serum-free medium containing sodium ion and calcium ion chelating agent when harvesting cells;
after the microcarrier is expanded and deformed, collecting cells in a physical mode;
wherein the microcarrier is the microcarrier or the microcarrier is the microcarrier obtained by the preparation method.
Preferably, the microcarrier and seed cell co-culture process needs periodic shaking to promote the adhesion of the seed cell to the microcarrier surface.
Further preferably, the culture process after replacement of the serum-free medium containing the sodium ion and calcium ion chelating agent requires periodic shaking to promote uniform suspension of the microcarriers to which the cells are adhered in the medium.
Specifically, the serum-free culture medium containing the sodium ion and calcium ion chelating agent comprises 100mmol/L sodium bicarbonate and 55mmol/L sodium citrate.
Optionally, the physically collecting cells comprises at least one of:
promoting the cells to be separated from the surface of the microcarrier by shaking or stirring;
separating the microcarriers and cells by a screen;
cells were enriched by centrifugation.
Preferably, the stirring speed is less than or equal to 100 r/min.
Alternatively, the seed cells are anchorage-dependent cultured cells.
Compared with the prior art, the method has the following advantages:
1. the main component of the microcarrier in the application contains calcium alginate composite. The material has the greatest advantage that in the presence of a calcium ion chelating agent (such as phosphate radical, carbonate radical, bicarbonate radical, citrate radical, EDTA and the like), the material is easy to expand and deform, so that the contact area between the material and cells is reduced, the cells can fall off under slight physical shearing force, and the separation of the cells and microcarriers is realized. The whole process can realize cell separation without the participation of pancreatin.
2. The microcarrier material is simultaneously compounded with a plurality of extracellular matrix materials to provide adhesion sites for cells, so that the cells can be adhered and proliferated on the surface of the microcarrier.
Drawings
FIG. 1 is a flow chart of a method of preparing a microcarrier according to the present application.
FIG. 2 is a flow chart of the cell mass culture method of the present application.
Detailed Description
The present application is described in further detail below with reference to the attached drawings and the detailed description.
The microcarriers of the present application consist essentially of gel molecules and extracellular matrix molecules. The gel molecules mainly refer to components that are cross-linked and polymerized to form a gel-like substance through a gelation reaction, and in this embodiment, alginate is preferably used, and specifically, sodium alginate and calcium alginate may be used. The extracellular matrix molecules mainly refer to components which can simulate the extracellular matrix, such as collagen, matrigel, chondroitin sulfate, fibronectin and chitosan, and can be single components or composite components according to the types of cultured cells; the presence of the extracellular matrix molecules allows cells to readily adhere to the surface of the microcarriers for proliferation. The microcarrier is easy to expand and deform in the presence of the calcium ion chelating agent, the contact area between the surface of the microcarrier subjected to expansion and deformation and cells is reduced, and the cells are easy to fall off from the surface of the microcarrier.
The preparation method of the microcarrier of the application is as follows:
respectively preparing a gel molecule solution and an outer matrix molecule solution;
mixing calcium carbonate powder with the gel molecular solution, and then mixing with the outer matrix molecular solution to form a premixed solution;
promoting the pre-mixed liquid to carry out gelation reaction by a fluid granulation technology to generate a hydrogel microcarrier;
and drying and sterilizing the hydrogel microcarrier to obtain dry powder of the microcarrier.
Further, the concentration of the gel molecule solution and the outer matrix solution affects the size of the hydrogel microcarrier, and a person skilled in the art can determine a specific value according to the experimental result, but preferably, the concentration of the gel molecule solution in the premix is kept above 10g/L to ensure that the gelation reaction is smoothly performed. The fluid granulation technology can be selected from high-voltage electrostatic field technology, gas orifice spraying technology, micro-fluidic technology or emulsification technology, and the emulsification technology can be further subdivided into emulsification-external gelation method, emulsification-internal gelation method and film emulsification method.
The preparation of microcarriers and the use of microcarriers in cell-scale cultivation methods according to the present application are further described below by way of specific examples.
Example one
Referring to fig. 1, the microcarrier is prepared as follows:
A. solution preparation: sodium alginate solution with concentration of 10-50 g/L; chitosan solution with the concentration of 1-10 g/L;
B. preparing a collagen solution, a chitosan solution, a matrigel solution, a chondroitin sulfate solution or a fibronectin solution with the concentration of 2-50 g/L;
C. mixing the sodium alginate solution prepared in the step A with calcium carbonate powder to prepare uniform suspension, and uniformly mixing the suspension with one or more of the collagen solution, the chitosan solution, the matrigel solution, the chondroitin sulfate solution or the fibronectin solution prepared in the step B, wherein the concentration of the sodium alginate in the mixed solution is kept above 10 g/L;
D. c, forming uniform liquid drops by the mixed liquid prepared in the step C through a fluid granulation technology, and carrying out a gelation reaction by calcium ion crosslinking and temperature change to obtain a spherical hydrogel microcarrier;
E. the spherical hydrogel microcarrier obtained in the step D can also be used as a core to further react with the chitosan solution prepared in the step A with the concentration of 1-10g/L, the volume ratio of the carrier to the chitosan solution is 1:2-1:10, the microcarrier is in a uniform suspension state in the chitosan solution, polyelectrolyte complex reaction is carried out for 2-60 minutes, namely a hydrogel film is formed on the surface of the core of the spherical hydrogel microcarrier, and the spherical hydrogel carrier with a core-shell structure is obtained after the physiological saline solution is cleaned;
F. and D, implanting the spherical hydrogel microcarrier prepared in the step D or the step E into an acid environment, and fully reacting with calcium carbonate powder in the carrier to generate carbon dioxide to generate the hydrogel microcarrier with a pore structure, wherein the pore structure of the hydrogel microcarrier can increase the surface area of the microcarrier, is favorable for increasing the cell adhesion area, and is also favorable for improving the material exchange efficiency in the hydrogel microcarrier.
G. And F, drying the spherical hydrogel carrier obtained in the step F in vacuum to obtain a dried microcarrier, and sterilizing the microcarrier by using high-pressure steam to obtain microcarrier dry powder for later use.
Example two
【1】 Preparing a sodium alginate solution with the concentration of 30 g/L;
【2】 Preparing a collagen solution with the concentration of 5g/L, and adjusting the concentration to be neutral by using NaOH;
【3】 Preparing a gel bath solution, wherein the gel bath contains calcium chloride (the concentration is 20 g/L);
【4】 Mixing the sodium alginate solution prepared in the step (1) with the collagen solution prepared in the step (2) in a volume ratio of 2: 1;
【5】 Forming stable jet flow by the mixed solution of the sodium alginate and the collagen prepared in the step (4) under a high-voltage electrostatic field, spraying the stable jet flow into the gel bath prepared in the step (3), and carrying out gelation reaction for 30 minutes at 37 ℃ in the gel bath to obtain the sodium alginate-collagen blended hydrogel microspheres;
【6】 And (5) sterilizing the sodium alginate-collagen blended hydrogel microspheres prepared in the step (5) under high pressure, and drying the sodium alginate-collagen blended hydrogel microspheres into microcarrier dry powder by a fluidized bed dryer for later use.
EXAMPLE III
A. The microcarrier obtained in the first embodiment or the second embodiment is cultured together with the cells to be cultured, and the cells to be cultured are uniformly adhered to the surface of the microcarrier by periodic shaking.
B. Preparing a serum-free medium containing specific ions: the serum-free medium contains sodium ions and one or more anions selected from phosphate radical, carbonate radical, bicarbonate radical, citrate radical and EDTA.
C. And D, culturing the microcarrier loaded with the cells obtained in the step A in a bioreactor, replacing the culture solution with the serum-free culture medium containing the sodium ion and calcium ion chelating agent prepared in the step B after the cells are expanded to a satisfactory degree, and periodically shaking after the culture solution is completely replaced to enable the microcarrier to be uniformly suspended in the solution prepared in the step B.
D. The microcarrier is expanded and deformed, cells are shed from the surface of the carrier into the solution under slight shearing force, the cells are physically separated through a screen with the aperture of 100 microns, the microcarrier is intercepted, and the cells in the supernatant are collected by centrifugation.
The process of the cell scale culture method is shown in FIG. 2, and the culture method is suitable for all anchorage-dependent cultured cells.
Example four
Taking the liver cell line HepG2 as an example, the cell scale culture method is used for culturing. The process is as follows:
the liver cell line HepG2 was inoculated onto microcarriers and cultured using a bioreactor.
Preparing a serum-free medium containing specific ions: contains 100mmol/L sodium bicarbonate and 55mmol/L sodium citrate.
Culturing liver cells on the microcarrier for 7 days, collecting the microcarrier, cleaning with normal saline, transferring to the serum-free culture medium containing specific ions prepared in the previous step, culturing in a bioreactor for 10 hours, stirring at the speed of 50 r/min, and collecting the cells after the microcarrier expands and the cells automatically fall off.
And (3) intercepting the expanded microcarrier by using a screen with the particle size of 100 microns in the mixture in the bioreactor, and centrifuging and collecting the cell suspension to obtain a cell product.
In summary, the cell mass culture method and the microcarrier used in the method are that cells are cultured on the surface of a microcarrier prepared from an extracellular matrix component such as sodium alginate and chitosan, the microcarrier and the cells are separated after the cells are cultured in a reactor for a certain time, specifically, a serum-free culture medium containing a sodium ion chelating agent and a calcium ion chelating agent is added into the microcarrier to expand the microcarrier, the adhesion degree of the cells and the surface of the microcarrier is reduced, the cells are separated, the expanded microcarrier is filtered and trapped by a screen, and the filtrate containing the cells is centrifuged to collect the cells.
The above embodiments are only preferred embodiments of the present application, but not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present application should be construed as equivalents and are included in the scope of the present application.
Claims (17)
1. A microcarrier comprising a gel molecule and an extracellular matrix molecule, wherein the gel molecule and the extracellular matrix molecule are mixed and then induced to undergo a gelation reaction to produce a hydrogel microcarrier; wherein the gel molecule is sodium alginate or calcium alginate, and the outer matrix molecule is one or more of collagen, matrigel protein, chondroitin sulfate, fibronectin and chitosan.
2. The microcarrier of claim 1, wherein the hydrogel microcarrier swells and deforms when soaked in a solution comprising a chelating agent for sodium and calcium ions.
3. The microcarrier of claim 2, wherein the calcium chelating agent is any one of phosphate, carbonate, bicarbonate, citrate, or EDTA.
4. A method for producing a microcarrier according to any one of claims 1 to 3, comprising the steps of:
respectively preparing a gel molecule solution and an outer matrix molecule solution;
mixing calcium carbonate powder with the gel molecular solution, and then mixing with the outer matrix molecular solution to form a premixed solution;
promoting the pre-mixed liquid to carry out gelation reaction by a fluid granulation technology to generate a hydrogel microcarrier;
and drying and sterilizing the hydrogel microcarrier to obtain dry powder of the microcarrier.
5. The method according to claim 4, wherein the concentration of the gel molecule solution is 10 to 50g/L, and the concentration of the outer matrix molecule solution is 1 to 50 g/L.
6. The method according to claim 4, wherein the concentration of the gel molecule solution in the premix is 10g/L or more.
7. The method of claim 4, wherein the fluid granulation technique is any one of high voltage electrostatic field technique, gas orifice spray technique, micro-fluidic technique, or emulsification technique.
8. The preparation method according to claim 4, wherein the hydrogel microcarrier is prepared by adding 1-10g/L of chitosan solution to perform polyelectrolyte complexation reaction to obtain the hydrogel microcarrier with a core-shell structure.
9. The method of claim 8, wherein the volume ratio of hydrogel microcarrier to chitosan solution is 1: 2-10; the duration time of the polyelectrolyte complexation reaction is 2-60 minutes.
10. The method of claim 4 or 8, wherein after the hydrogel microcarrier is formed, the hydrogel microcarrier is placed in an acidic environment to produce carbon dioxide, thereby forming a hydrogel microcarrier having a porous structure.
11. A cell scale culture method is characterized by comprising the following steps:
preparing microcarrier and seed cell separately;
co-culturing the microcarrier and the seed cells by using a serum-free culture medium which does not contain a sodium ion and calcium ion chelating agent, so that the seed cells are adhered to the surface of the microcarrier for proliferation;
replacing serum-free medium containing sodium ion and calcium ion chelating agent when harvesting cells;
after the microcarrier is expanded and deformed, collecting cells in a physical mode;
wherein the microcarrier is the microcarrier according to any one of claims 1 to 3, or the microcarrier is the microcarrier obtained by the preparation method according to any one of claims 4 to 10.
12. The culture method according to claim 11, wherein the co-culture of the microcarrier and the seed cell requires periodic shaking to promote the adhesion of the seed cell to the microcarrier surface.
13. The culture method according to claim 11, wherein the culture process after replacing the serum-free medium containing the sodium ion and calcium ion chelating agent requires periodic shaking to promote uniform suspension of the microcarriers to which the cells are adhered in the medium.
14. The culture method according to claim 11, wherein the serum-free medium containing the sodium ion-and calcium ion-chelating agent comprises 100mmol/L sodium bicarbonate and 55mmol/L sodium citrate.
15. The culture method of claim 11, wherein physically collecting the cells comprises at least one of:
promoting the cells to be separated from the surface of the microcarrier by shaking or stirring;
separating the microcarriers and cells by a screen;
cells were enriched by centrifugation.
16. The culture method according to claim 15, wherein the stirring speed is 100r/min or less.
17. The culture method of claim 11, wherein the seed cells are anchorage-dependent cultured cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110321555.6A CN113061566A (en) | 2021-03-25 | 2021-03-25 | Cell large-scale culture method and microcarrier used by same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110321555.6A CN113061566A (en) | 2021-03-25 | 2021-03-25 | Cell large-scale culture method and microcarrier used by same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113061566A true CN113061566A (en) | 2021-07-02 |
Family
ID=76563620
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110321555.6A Pending CN113061566A (en) | 2021-03-25 | 2021-03-25 | Cell large-scale culture method and microcarrier used by same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113061566A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116103218A (en) * | 2023-02-03 | 2023-05-12 | 国纳科技(昆山)有限公司 | Preparation method and product of 3D cell culture hydrogel microsphere |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004033135A (en) * | 2002-07-05 | 2004-02-05 | Fuji Photo Film Co Ltd | Carrier for culturing cell |
| US20040072338A1 (en) * | 2002-07-05 | 2004-04-15 | Fuji Photo Film Co., Ltd. | Carrier for cell culture |
| CN102475691A (en) * | 2010-11-30 | 2012-05-30 | 中国科学院大连化学物理研究所 | Alginate-chitosan acyl derivative microcapsules and its preparation and application |
| CN105087465A (en) * | 2015-08-26 | 2015-11-25 | 南方医科大学珠江医院 | Hepatocyte serum-free culture medium |
| CN107304409A (en) * | 2015-12-11 | 2017-10-31 | 香港城市大学 | substrate for three-dimensional cell culture, preparation method and application thereof |
| CN107961228A (en) * | 2018-01-25 | 2018-04-27 | 南方医科大学 | A kind of sodium alginate-chitosan P-VP8*IgY microcapsule preparation methods and microcapsules |
-
2021
- 2021-03-25 CN CN202110321555.6A patent/CN113061566A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004033135A (en) * | 2002-07-05 | 2004-02-05 | Fuji Photo Film Co Ltd | Carrier for culturing cell |
| US20040072338A1 (en) * | 2002-07-05 | 2004-04-15 | Fuji Photo Film Co., Ltd. | Carrier for cell culture |
| CN102475691A (en) * | 2010-11-30 | 2012-05-30 | 中国科学院大连化学物理研究所 | Alginate-chitosan acyl derivative microcapsules and its preparation and application |
| WO2012072012A1 (en) * | 2010-11-30 | 2012-06-07 | 中国科学院大连化学物理研究所 | Microcapsule preparation of alginate-chitosan acyl derivatives, preparation and application thereof |
| CN105087465A (en) * | 2015-08-26 | 2015-11-25 | 南方医科大学珠江医院 | Hepatocyte serum-free culture medium |
| CN107304409A (en) * | 2015-12-11 | 2017-10-31 | 香港城市大学 | substrate for three-dimensional cell culture, preparation method and application thereof |
| CN107961228A (en) * | 2018-01-25 | 2018-04-27 | 南方医科大学 | A kind of sodium alginate-chitosan P-VP8*IgY microcapsule preparation methods and microcapsules |
Non-Patent Citations (2)
| Title |
|---|
| SONG YIZHE 等: "Microfabrication of a tunable collagen/alginate-chitosan hydrogel membrane for controlling cell-cell interactions", 《CARBOHYDR POLYM.》 * |
| 宋益哲 等: "胶原／海藻酸钙互穿网络水凝胶的构建及其对细胞行为的影响", 《功能材料》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116103218A (en) * | 2023-02-03 | 2023-05-12 | 国纳科技(昆山)有限公司 | Preparation method and product of 3D cell culture hydrogel microsphere |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110272860B (en) | Construction method and application of three-dimensional cell culture microenvironment | |
| Bölgen et al. | Three-dimensional ingrowth of bone cells within biodegradable cryogel scaffolds in bioreactors at different regimes | |
| US9926528B2 (en) | Glucomannan scaffolding for three-dimensional tissue culture and engineering | |
| WO2008005520A2 (en) | Temperature-responsive microcarrier | |
| CN111330082B (en) | Preparation method for constructing biological 3D printing skin micro-unit model containing skin accessory | |
| Prakash et al. | Preparation and in vitro analysis of microencapsulated genetically engineered E. coli DH5 cells for urea and ammonia removal | |
| JP2016523086A (en) | Cell culture article and method thereof | |
| CN114317394A (en) | Microcarrier for three-dimensional cell culture and preparation method and application thereof | |
| Mahboubian et al. | Temperature-responsive methylcellulose–hyaluronic hydrogel as a 3D cell culture matrix | |
| CN111676191A (en) | Method for three-dimensionally culturing mesenchymal stem cells based on Collagel gel scaffold method | |
| CN113061566A (en) | Cell large-scale culture method and microcarrier used by same | |
| Li et al. | Preparation of microcarriers based on zein and their application in cell culture | |
| KR20230081976A (en) | Hybrid cultured meat comprising grains and a cultured cell coating layer located on the surface of the grains and method for thereof | |
| Sakai et al. | Wrapping tissues with a pre-established cage-like layer composed of living cells | |
| US20120190113A1 (en) | Macroporous Microcarrier Specific to Liver Cell, Preparation Method and Use Thereof | |
| CN105734017B (en) | A method for promoting the directed differentiation and proliferation of mesenchymal stem cells to neural precursor cells | |
| CN108102915A (en) | A kind of mediate contact co-culture system for being engineered amplification | |
| CN111705036A (en) | Method for three-dimensionally culturing fiber ring source stem cells based on Collagel gel scaffold method | |
| JP2628536B2 (en) | Cell culture substrate | |
| CN120187843A (en) | Dissolvable foam scaffold for cell culture and method of making same | |
| CN113318270B (en) | Method for preparing bioactive substance coated polyester mesh lamellar stent | |
| Zhou et al. | Microspheres for cell culture | |
| JPH06277050A (en) | Immobilization material for animal cell and culture method | |
| CN116209747A (en) | Activated soluble carrier for affinity binding and cell culture | |
| CN111876379A (en) | Method for efficiently promoting proliferation of fiber ring source stem cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210702 |