CN107012115A - Culture medium of sertoli cell high density suspension culture and preparation method thereof - Google Patents
Culture medium of sertoli cell high density suspension culture and preparation method thereof Download PDFInfo
- Publication number
- CN107012115A CN107012115A CN201710255980.3A CN201710255980A CN107012115A CN 107012115 A CN107012115 A CN 107012115A CN 201710255980 A CN201710255980 A CN 201710255980A CN 107012115 A CN107012115 A CN 107012115A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- culture
- high density
- sertoli cell
- cell high
- 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
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
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0681—Cells of the genital tract; Non-germinal cells from gonads
- C12N5/0682—Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells
-
- 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
-
- 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
- C12N2500/05—Inorganic components
- C12N2500/10—Metals; Metal chelators
- C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
- C12N2500/14—Calcium; Ca chelators; Calcitonin
-
- 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
- C12N2500/20—Transition metals
-
- 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
- C12N2500/20—Transition metals
- C12N2500/22—Zinc; Zn chelators
-
- 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
- C12N2500/20—Transition metals
- C12N2500/24—Iron; Fe chelators; Transferrin
-
- 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
- C12N2500/20—Transition metals
- C12N2500/24—Iron; Fe chelators; Transferrin
- C12N2500/25—Insulin-transferrin; Insulin-transferrin-selenium
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/32—Amino acids
-
- 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
- C12N2500/34—Sugars
-
- 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
- C12N2500/38—Vitamins
-
- 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
- C12N2500/40—Nucleotides, nucleosides, bases
-
- 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
- C12N2500/46—Amines, e.g. putrescine
-
- 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
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
- C12N2500/95—Protein-free medium and culture conditions
Abstract
The present invention relates to a kind of culture medium of sertoli cell high density suspension culture, the culture medium contains several amino acids, inorganic salts, trace element, vitamin and other additives, without protein, its specific chemical components, component is stable between culture medium batch, it is cheap, it can solve the problem that the unstable adverse effect caused of nutrient media components in cell heavy industrialization incubation.
Description
Technical field
The present invention relates to field of cell culture, the culture medium of more particularly to a kind of sertoli cell high density suspension culture and
Its preparation method.
Background technology
Cell culture medium is divided into basal medium and serum free medium from occupation mode.Basal medium main component
Have：Inorganic salts, amino acid, carbon source, vitamin, buffer system and other class trace nutrients, its definite ingredients.But,
Basal medium can not fully meet the growth of cell, breed, it is necessary to coordinate serum to use.The main function of serum is to thin
Born of the same parents provide hormone, growth factor, transfer protein and the other nutriments needed for growing multiplication, and its ingredient is complicated, and not
There is mass discrepancy between the same place of production, batch, thus many adverse effects are caused to zooblast large-scale culture technique.Example
Such as, during obtaining product by cell culture, serum is the major obstacle of purifying, and it can even make product be difficult to turn into doctor
Medicine product.
In the production of industrial-scale metaplasia, it is thin that conventional cell line includes Chinese hamster ovary celI, Vero cells, SP2/0 cells, NS0
Born of the same parents, mouse hybridoma cell etc., most common of which are Chinese hamster ovary celIs.Up to now, 63 of U.S. FDA approval have listed
In antibody drug expression system, there are 33 using Chinese hamster ovary celI, occupy 52%.The expression of Chinese hamster ovary celI intrinsic protein is low, expression production
Thing is directly secreted into culture medium, and product is easy to collect.Readily suspended culture, is especially suitable for industrializing large-scale culture.With
Raising of the people to biological products security requirement, serum free medium, or even chemical composition limit protein-free medium by
Gradually turn into development trend from now on.
The protein-free medium that Chinese hamster ovary celI chemical composition domestic at present is limited rely primarily on gibico, sigma,
The import of Reagent Company of the foreign countries such as hyclone, longza.The country devises many serum free mediums also for Chinese hamster ovary celI, mainly
Method be on the basis of basal medium such as DMEM/F12, add protolysate, insulin, transferrins, growth factor,
Vitamin, lipid, trace element etc., these all albumen or vegetable protein hydrolyzate containing animal origin to a certain extent,
Risk and destabilizing factor are brought to bio-pharmaceuticals.Meanwhile, all kinds of recombinant proteins are expensive to cause culture medium cost expensive, pushes away
The high cost of whole technique.Therefore, the serum free medium that exploitation Chinese hamster ovary celI chemical composition is limited is significant.
The content of the invention
Based on this, it is necessary in view of the above-mentioned problems, it is high to provide a kind of specific chemical components, nonprotein sertoli cell
Culture medium of density suspension culture and preparation method thereof.
To achieve these goals, the present invention uses following technical scheme：
A kind of culture medium of sertoli cell high density suspension culture, including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
In wherein some embodiments, the chemical composition defined medium of the support Chinese hamster ovary celI high density suspension culture,
Including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
In wherein some embodiments, the culture medium of the sertoli cell high density suspension culture, including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
In wherein some embodiments, the cell for high density suspension culture is Chinese hamster ovary celI.
The present invention also provides a kind of preparation method of the culture medium of sertoli cell high density suspension culture, including following step
Suddenly：
(1) mixed solution is prepared：By dissolution of raw material in a solvent, mixed solution is obtained；
(2) fluid nutrient medium is prepared：By mixed liquor filtration sterilization, fluid nutrient medium is obtained.
In wherein some embodiments, solvent described in step (1) is without thermal source aqua sterilisa.
In wherein some embodiments, in step (1), after dissolution of raw material, pH is to 7.2-7.5 for regulation, obtains mixed solution.
In culture medium of the present invention, transferrins is substituted using aurin tricarboxyli acid (ATA), ferric sulfate by optimum organization, adopted
Insulin is substituted with nickel chloride and caddy, common combination obtains the serum free medium of the present invention, and it is free of albumen
Component is stable between matter, specific chemical components, culture medium batch, cheap, can solve the problem that cell heavy industrialization culture
During the unstable adverse effect caused of nutrient media components, it can also well support Chinese hamster ovary celI in vitro high density hang
Floating culture, meets Chinese hamster ovary celI culture demand.
Brief description of the drawings
Fig. 1 is growth curve of the CHO-k1 cells in different culture media.
Fig. 2 is Cell viability of the CHO-k1 cells in different culture media.
Fig. 3 is the displaing micro picture of the adherent type CHO-k1 cell lines before the domestication of embodiment 1.
Fig. 4 is the displaing micro picture of the serum free suspension type CHO-k1 cell lines after the domestication of embodiment 1.
Embodiment
For the objects, technical solutions and advantages of the present invention are more clearly understood, below in conjunction with embodiment, to this
Invention is described in further detail.It should be appreciated that embodiment described herein is only to explain this hair
It is bright, do not limit protection scope of the present invention.
1st, the culture medium of sertoli cell high density suspension culture is prepared
The nutrient media components and consumption of embodiment 1-3 sertoli cell high density suspension culture are as shown in the table：
Each component is sigma cell culture grade raw materials in above-mentioned table, and prepares culture medium according to following methods：
The each component raw material of embodiment 1, embodiment 2 and embodiment 3 is added separately in 800ml distilled waters, 5M is used
NaCl regulation infiltrations are depressed into 270-275mosm, pH to 7.2-7.5 are adjusted with 1N HCl or NaOH solution, then with 0.22 μm
Membrane filtration is degerming, 4 DEG C of preservations, obtains the culture medium cultivated that suspended for high cell densities.
2nd, cell culture
It is used for the culture medium of high cell densities suspension culture described in Application Example 1, cell is cultivated, meanwhile,
Using commercially available sigma CD-CHO culture mediums and gibico CD-CHO culture mediums as control, method is as follows：
(1) the CHO-k1 cells adapted to will be suspended with 5 × 105Individual/mL density is seeded to the 125mL of the culture medium containing 20mL
In shaking flask, shaking flask is placed in 37 DEG C of cultures in the shaking table containing 5% carbon dioxide, rotating speed is 125rpm, passed on once within every 2 days, half
Amount changes liquid and carries out Secondary Culture, and continuous passage 3 is more than generation with acclimatizing culture medium.
(2) by the cell in exponential phase in (1) with 5 × 105Individual/mL density is inoculated into the culture medium containing 20mL
In 125mL shaking flasks, shaking flask is placed in 37 DEG C of cultures in the shaking table containing 5% carbon dioxide, rotating speed is 125rpm, continuous culture 7 days,
Count daily, and with Trypan Blue meter Cell viability.
3rd, experimental result
Referring to Fig. 1, the 4th day after cell inoculation, cell density reaches highest.Culture medium described in embodiments of the invention 1,
Commercially available sigma culture medium and commercially available gibico culture medium cell density respectively reach 6 × 106Individual/ml, 5.5 × 106Individual/ml
With 5 × 106Individual/ml.Cell viability was held at more than 95%, the accumulation of later stage nutrient consumption and metabolic waste at first 4 days
Cell viability is caused to be gradually reduced, three kinds of culture medium difference are little, referring to Fig. 2.In general, reality of the Chinese hamster ovary celI in the present invention
The growing state applied in culture medium described in example 1 is better than import culture medium.Sigma serum-frees are slightly better than gibico free serum culture
Base, it may be possible to which sigma serum free medium is more suitable for the cell line.
From Fig. 3 and Fig. 4, CHO-k1 cells cell attachment growth before domestication of the embodiment of the present invention 1, in shuttle
Shape.The serum free suspension type CHO-k1 cells obtained after domestication are spherical in shape, and good dispersion, the bright growth conditions of cell are preferable.
The experimental result of embodiment 1 see the table below：
It is used for the culture medium of high cell densities suspension culture described in Application Example 2 and embodiment 3, cell is trained
Support, method is as follows：
(1) the CHO-k1 cells adapted to will be suspended with 5 × 105Individual/mL density is seeded to embodiment containing 20mL 2 or implementation
In the 125mL shaking flasks of the culture medium of example 3, shaking flask is placed in 37 DEG C of cultures in the shaking table containing 5% carbon dioxide, rotating speed is 125rpm,
Once, half amount changes liquid and carries out Secondary Culture, and continuous passage 3 is more than generation with acclimatizing culture medium for passage in every 2 days.
(2) by the cell in exponential phase in (1) with 5 × 105Individual/mL density is inoculated into embodiment containing 20mL 2
Or in the 125mL shaking flasks of the culture medium of embodiment 3, shaking flask is placed in 37 DEG C of cultures in the shaking table containing 5% carbon dioxide, rotating speed is
125rpm, continuous culture 7 days, is counted daily, and with Trypan Blue meter Cell viability.
The experimental result of embodiment 2 see the table below：
The experimental result of embodiment 3 see the table below：
Test result indicates that, embodiment 2 and example 3 are equally supported in the normal growth of Chinese hamster ovary celI, embodiment 2, each nutrition into
Point content is slightly lower, and cell growth rate and survival rate are relatively low；In embodiment 3, each nutrient composition content is higher, cell growth rate
It is all higher with survival rate, but be more or less the same with embodiment 1, consider, each component content of embodiment 1 is most saved, effect also compared with
It is good, it is most economical combination.
Embodiment 4
, it is necessary to avoid adding all kinds of recombinant proteins and chemical composition is not clear in the serum free medium that chemical composition is limited
True composition.The absorption that addition transferrins promotes iron is generally required in traditional serum free medium, adds insulin to promote
The absorption of glucose.The present invention adds aurin tricarboxyli acid (ATA), ferric sulfate to substitute transferrins, adds nickel chloride, caddy to replace
For insulin, with reference to other components, recombinant protein and animal derived components are effectively substituted for, and reduce the cost of culture medium.
In example 4, using in the recipe ingredient of embodiment 1 remove aurin tricarboxyli acid (ATA), ferric sulfate, nickel chloride, caddy as
Group A, culture medium described in Application Example 1 is group B, and transferrins (5.5mg/L) and pancreas islet are added on the basis of group A culture mediums
Plain (10mg/L) is that the culture medium after addition nickel chloride, caddy is a group D on the basis of a group C, group A culture mediums, group A culture mediums
On the basis of addition insulin (10mg/L) be a group E, the training on the basis of group A culture mediums after addition aurin tricarboxyli acid (ATA) and ferric sulfate
It is group F to support base, and it is group G that transferrins (5.5mg/L) is added on the basis of group A culture mediums.Transferrins and insulin are purchased
From sigma.The CHO-k1 cells cultivated are suspended as seed cell using the adaptation in growth logarithmic phase, with the first of 5 × 10e5/ml
Beginning density is inoculated with, continuous culture 3 days, counts viable count.As a result it is as follows：
Group A vitro growth rates without aurin tricarboxyli acid (ATA), ferric sulfate, nickel chloride, caddy are decreased obviously, and cell occurs
Agglomerate.And the cell growth rate containing aurin tricarboxyli acid (ATA), ferric sulfate, nickel chloride, the group B of caddy is higher than containing insulin and turned
The replacement (D/E, F/G group) of the group C of ferritin, insulin and transferrins independent component all shows close culture effect,
Show its superior substitution effect.
Each technical characteristic of embodiment described above can be combined arbitrarily, to make description succinct, not to above-mentioned reality
Apply all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, the scope of this specification record is all considered to be.
Embodiment described above only expresses the several embodiments of the present invention, and it describes more specific and detailed, but simultaneously
Can not therefore it be construed as limiting the scope of the patent.It should be pointed out that coming for one of ordinary skill in the art
Say, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (7)
1. a kind of culture medium of sertoli cell high density suspension culture, it is characterised in that including following components：Inorganic salts with it is micro
Element：
Vitamin：
Amino acid：
Other additives：
2. the culture medium of sertoli cell high density suspension culture according to claim 1, it is characterised in that including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
3. the culture medium of sertoli cell high density suspension culture according to claim 2, it is characterised in that including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
4. the culture medium of sertoli cell high density suspension culture according to claim 3, it is characterised in that including following components：
Inorganic salts and trace element：
Vitamin：
Amino acid：
Other additives：
5. a kind of method for the culture medium for preparing any described sertoli cell high density suspension cultures of claim 1-4, it is special
Levy and be, comprise the following steps：
(1) mixed solution is prepared：By dissolution of raw material in a solvent, mixed solution is obtained；
(2) fluid nutrient medium is prepared：By mixed liquor filtration sterilization, fluid nutrient medium is obtained.
6. the preparation method of the culture medium of sertoli cell high density suspension culture according to claim 5, it is characterised in that
Solvent described in step (1) is without thermal source aqua sterilisa.
7. the preparation method of the culture medium of sertoli cell high density suspension culture according to claim 5, it is characterised in that
In step (1), after dissolution of raw material, pH is to 7.2-7.5 for regulation, obtains mixed solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710255980.3A CN107012115A (en) | 2017-04-18 | 2017-04-18 | Culture medium of sertoli cell high density suspension culture and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710255980.3A CN107012115A (en) | 2017-04-18 | 2017-04-18 | Culture medium of sertoli cell high density suspension culture and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107012115A true CN107012115A (en) | 2017-08-04 |
Family
ID=59447613
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710255980.3A Pending CN107012115A (en) | 2017-04-18 | 2017-04-18 | Culture medium of sertoli cell high density suspension culture and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107012115A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108624550A (en) * | 2018-02-14 | 2018-10-09 | 世贸天阶制药（江苏）有限责任公司 | The additive and culture medium obtained of a kind of Chinese hamster ovary celI serum free medium |
| CN110835622A (en) * | 2018-08-16 | 2020-02-25 | 上海药明生物技术有限公司 | Culture medium for regulating lactic acid metabolism of mammalian cells and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1867665A (en) * | 2003-08-08 | 2006-11-22 | 剑桥抗体技术有限公司 | Myeloma cell culture in transferrin-free low iron medium |
| CN101724600A (en) * | 2009-11-30 | 2010-06-09 | 中国人民解放军军事医学科学院生物工程研究所 | Blood serum-free culture medium for supporting suspension culture of CHO cells with large scale and high density |
| CN102443565A (en) * | 2010-09-30 | 2012-05-09 | 成都康弘生物科技有限公司 | Medium suitable for cultivating CHO cell and cultivation technology thereof |
| CN102876626A (en) * | 2011-07-14 | 2013-01-16 | 宁波安柯普顿生物技术有限公司 | Serum-free and protein-free all-chemical-component-definition culture medium for supporting CHO high-density suspension growth |
| CN103773732A (en) * | 2013-06-08 | 2014-05-07 | 李锋 | Chemically-defined medium, application thereof and production technology for large-scale culture of mammalian cells |
| CN104073463A (en) * | 2013-03-29 | 2014-10-01 | 上海中信国健药业股份有限公司 | Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture |
| CN104328158A (en) * | 2014-09-29 | 2015-02-04 | 华东理工大学 | Chemical-component-definite culture medium suitable for large-scale production of animal cell expression production |
-
2017
- 2017-04-18 CN CN201710255980.3A patent/CN107012115A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1867665A (en) * | 2003-08-08 | 2006-11-22 | 剑桥抗体技术有限公司 | Myeloma cell culture in transferrin-free low iron medium |
| CN101724600A (en) * | 2009-11-30 | 2010-06-09 | 中国人民解放军军事医学科学院生物工程研究所 | Blood serum-free culture medium for supporting suspension culture of CHO cells with large scale and high density |
| CN102443565A (en) * | 2010-09-30 | 2012-05-09 | 成都康弘生物科技有限公司 | Medium suitable for cultivating CHO cell and cultivation technology thereof |
| CN102876626A (en) * | 2011-07-14 | 2013-01-16 | 宁波安柯普顿生物技术有限公司 | Serum-free and protein-free all-chemical-component-definition culture medium for supporting CHO high-density suspension growth |
| CN104073463A (en) * | 2013-03-29 | 2014-10-01 | 上海中信国健药业股份有限公司 | Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture |
| CN103773732A (en) * | 2013-06-08 | 2014-05-07 | 李锋 | Chemically-defined medium, application thereof and production technology for large-scale culture of mammalian cells |
| CN104328158A (en) * | 2014-09-29 | 2015-02-04 | 华东理工大学 | Chemical-component-definite culture medium suitable for large-scale production of animal cell expression production |
Non-Patent Citations (1)
| Title |
|---|
| Y.M. QI ET AL.: ""Evaluation of a simple protein free medium that supports high levels of monoclonal antibody production"", 《CYTOTECHNOTOGY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108624550A (en) * | 2018-02-14 | 2018-10-09 | 世贸天阶制药（江苏）有限责任公司 | The additive and culture medium obtained of a kind of Chinese hamster ovary celI serum free medium |
| CN110835622A (en) * | 2018-08-16 | 2020-02-25 | 上海药明生物技术有限公司 | Culture medium for regulating lactic acid metabolism of mammalian cells and application thereof |
| CN110835622B (en) * | 2018-08-16 | 2021-04-27 | 上海药明生物技术有限公司 | Culture medium for regulating lactic acid metabolism of mammalian cells and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106987554B (en) | Suspension cell strain and domestication method thereof | |
| CN102827804B (en) | Culture medium and method suitable for the amplification culture of Vero cell microcarrier suspension | |
| CN101864393B (en) | Serum-free culture medium without animal origin components for culturing Vero cell micro-carrier | |
| Chun et al. | Usability of size-excluded fractions of soy protein hydrolysates for growth and viability of Chinese hamster ovary cells in protein-free suspension culture | |
| CN100348718C (en) | Culture medium without animal originating component and serum for HEK293 cell adhesion culture | |
| CN104560882B (en) | A kind of Chinese hamster ovary celI culture process for reducing acidic variants content | |
| CN103773732B (en) | The production technology of definite culture medium, its application and the large-scale culture mammalian cell of a kind of chemical composition | |
| CN103898041B (en) | The cultural method of hybridoma | |
| CN104164405A (en) | Serum-free culture system for efficiently culturing human umbilical cord mesenchymal stem cells in vitro | |
| CN102549142A (en) | Method of producing a polypeptide or virus of interest in a continuous cell culture | |
| CN106190950A (en) | A kind of Chinese hamster ovary celI Serum-free and protein-free medium and preparation method thereof | |
| CN109370985A (en) | A kind of human umbilical cord mesenchymal stem cells large-scale culture serum free medium | |
| CN107012115A (en) | Culture medium of sertoli cell high density suspension culture and preparation method thereof | |
| CN111454877B (en) | CHO cell culture method | |
| CN101245335A (en) | Non-blood serum plant protein peptide universal cell culture medium | |
| CN103146525B (en) | Production method of soft type composite multi-micro-function yeast | |
| CN109609444A (en) | A kind of BHK21Cell non-serum culture medium | |
| CN111748527B (en) | Chemical component limited efficient feeding culture medium and preparation method and application thereof | |
| WO2021008571A1 (en) | Cell culture method and application thereof based on high-density and continuous inoculation | |
| CN103484426B (en) | Non-animal source low-protein culture medium | |
| CN102405279B (en) | Improve the method for single cell clone | |
| CN109576212B (en) | Culture method of seed cells in high-density inoculation culture and application thereof | |
| CN107904200B (en) | It is a kind of expression adalimumab Combined culture base and its application | |
| CN104164404A (en) | Application of serum-free culture system for efficiently culturing human umbilical cord mesenchymal stem cells in vitro | |
| Hermes et al. | A fully defined, fed‐batch, recombinant NS0 culture process for monoclonal antibody production |
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: 20170804 |