CN104593316B - Insect cell serum free medium and its application - Google Patents
Insect cell serum free medium and its application Download PDFInfo
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- CN104593316B CN104593316B CN201510053687.XA CN201510053687A CN104593316B CN 104593316 B CN104593316 B CN 104593316B CN 201510053687 A CN201510053687 A CN 201510053687A CN 104593316 B CN104593316 B CN 104593316B
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Abstract
The invention discloses a kind of insect cell serum free medium and its application.The culture medium includes amino acid, inorganic salts, vitamin and carbohydrate etc.；Further, also comprising 0.5~15mg/L dilinoleoylphosphatidylcholines and/or 0.1~10mg/L DSPCs；Especially, 0.001~0.1mg/L barium chlorides, 0.005~0.02mg/L lithium chlorides, 0.05~7mg/L nickel chlorides can also be included.The insect cell serum free medium component of the present invention is simply clear and definite, and easily prepared, cost is cheap, and Insect cellculture efficiency and expression of recombinant proteins efficiency can be substantially improved, beneficial to the extensive preparation for the large-scale culture and recombinant protein for realizing insect cell.
Description
Technical field
The present invention relates to a kind of serum free medium, particularly one kind to be suitable to bioreactor large-scale culture Prepare restructuring
The insect cell serum free medium of albumen.
Background technology
To meet basic research and applying biomedical sector such as cell biology, physiology, pharmacology and toxicology day
Benefit is widely applied demand, and cell culture technology arises at the historic moment.Generally, the body of cell culture technology system analog cell growth
Interior environment, including temperature, pH value, osmotic pressure and oxygen supply, so that cell can grow and breed well.Wherein most critical
Factor is culture medium.
Conventional cell culture medium be mainly by basal medium add respective amount serum or tissue extract and
Formed, wherein because of the limitation of animal blood serum extraction, make it fairly expensive applied to the price of culture medium, and give cell culture
Standardization bring difficulty, while the problem of express separation and purification of products difficulty there is also cell culture, there is potential cell toxicant
Property effect, add difficulty to pilot scale culture cell.
Due to problems existing for traditional serum cell culture medium, researcher is promoted to develop free serum culture again
Base.Compared with traditional culture medium, serum free medium is free of animal blood serum or other biological extract solution, has constituent phase
To clear, the advantages that preparation process is simple, but still cell long period growth, breeding, and blood can be avoided in vitro can be maintained
Toxic action and serum source contact scar clearly to cell etc. adversely affect, and are advantageous to the differentiation of cultured cell in vitro, can improve production
The expression of product simultaneously makes cell products be easy to purify.Certainly, yet there are some defects in existing serum free medium, such as
Preserve and application is not so good as traditional synthetic media conveniently, cost is higher, and specific aim is very strong, and a kind of serum free medium is suitable only for
Culture of certain a kind of cell etc..
On the other hand, in genetic engineering field, increasingly it is taken seriously come express express target protein using eukaryotic expression system,
Conventional eukaryotic expression system has yeast expression system, insect cell expression system and mammalian cell expression system.Wherein
Due to its safe operation, expression quantity is high, gains great popularity at present for insect cell expression system, particularly baculovirus expression system,
And it has been widely used in producing exogenous proteins.
So far, such as L.Ikonomou., and etal (Appl Microbiol Biotechnol, 2003,62:1-20),
Michael S., etal (Biotechnol.Prog., 1988,14,573-579), RH Qi Erruiluo (CN1498267A)
Deng developed it is a variety of be used to cultivate the serum free mediums of insect cell, but these culture medium generally existing costs are too high,
Insect cellculture efficiency is low, and protein expression efficiency has the problems such as to be hoisted, is unfavorable for large-scale Insect cellculture and again
The preparation of histone.
The content of the invention
It is a primary object of the present invention to provide a kind of elder brother suitable for bioreactor large-scale culture Prepare restructuring albumen
Worm cell non-serum culture medium, to overcome deficiency of the prior art.
To realize aforementioned invention purpose, the technical solution adopted by the present invention includes：
A kind of insect cell serum free medium, include amino acid, inorganic salts, vitamin and carbohydrate, its feature
It is that it also includes 0.6~25mg/L phosphatidyl cholines, the phosphatidyl choline includes dilinoleoylphosphatidylcholine, distearyl
Phosphatidyl choline.
As more one of preferred embodiment, the insect cell medium includes 11200~29170mg/L amino
Acid, 5500.71~9510.33mg/L inorganic salts, 11.354~820.4mg/L vitamins, 3000~11000mg/L proteolysis
Thing and 6042~14222mg/L carbohydrate.
As one of embodiment particularly preferably, the insect cell medium includes 0.5~15mg/L, bis- sub- oleoyls
Phosphatidyl choline (Dilineoyl-phosphatidylcholin) and 0.1~10mg/L DSPCs
At least one of (Distearoyl-phosphatidylcholin), preferably use the rwo simultaneously.
As one of highly preferred embodiment, the insect cell medium also includes the water of 0.001~0.1mg/L bis-
Barium chloride, 0.005~0.02mg/L lithium chlorides, 0.05~7mg/L nickel chlorides.
As more one of preferred embodiment, the insect cell medium includes 300~800mg/ of Beta-alanine
L, 800~2400mg/L of L- R-genes, 900~1700mg/L of altheine, L-Aspartic acid sodium salt 1000~
1600mg/L, 150~300mg/L of CYSTINE hydrochloride, 1400~2700mg/L of Pidolidone hydrochloride, Glu
1000~5000mg/L, 300~800mg/L of glycine, 400~900mg/L of L-Histidine, 500~1500mg/ of L- hydroxyprolines
L, 500~1200mg/L of ILE, 200~600mg/L of L-Leu, 500~1500mg/L of LYS, L- eggs
700~1600mg/L of propylhomoserin, 800~1700mg/L of L-phenylalanine, 400~950mg/L of L-PROLINE, Serine 300~
1000mg/L, 300~820mg/L of L-threonine, 100~300mg/L of L-Trp, 300~850mg/ of TYR disodium salt
L, 350~950mg/L of Valine.
As more one of preferred embodiment, the insect cell medium include ammonium molybdate tetrahydrate 0.03~
0.2mg/L, 0.03~0.35mg/L of CoCL2 6H2O, 0.1~1.5mg/L of copper chloride dihydrate, ferrous sulfate heptahydrate 0.5~
4.6mg/L, 500~1500mg/L of anhydrous magnesium sulfate, 0.01~0.18mg/L of tetrahydrate manganese chloride, 1000~1500mg/ of potassium chloride
L, 3000~5000mg/L of sodium chloride, 1000~1500mg/L of sodium dihydrogen phosphate-water, 0.04~3.5mg/L of zinc chloride.
As more one of preferred embodiment, the insect cell medium include 0.1~0.5mg/L of biotin,
0.004~0.2mg/L of D-VB5 calcium, 10~400mg/L of Choline Chloride, 0.2~1.6mg/L of VB12 cyanocobalamins, VB9 folic acid 0.05
0.2~400mg/L of~0.5mg/L, i- inositols, 0.1~1.9mg/L of VB3 nicotinic acid, 0.3~2.7mg/L of p-aminobenzoic acid, VB6
0.3~4mg/L of puridoxine hydrochloride, 0.05~5mg/L of VB2 riboflavin, 0.05~4mg/L of VB1 thiamine hydrochlorides.
As more one of preferred embodiment, the insect cell medium include DEXTROSE ANHYDROUS 2500~
5000mg/L, 2~8mg/L of fumaric acid, 10~40mg/L of 2-oxoglutaric acid, 10~100mg/L of L MALIC ACID, a water Fructus Hordei Germinatus
Sugar (ultrapure) 500~3000mg/L, 4~14mg/L of succinic acid, Twen8010~1000mg/L, Cholesterol sulfate ester sodium salt 5~
1~10mg/L of 50mg/L, VE sodium, 3000~5000mg/L of D- sucrose.
As more one of preferred embodiment, the insect cell medium also includes protolysate, such as can
Preferably comprise 1000~5000mg/L of barley hydrolysate, 2000~6000mg/L of rice hydrolysate.
Among of the invention one more specifically embodiment, the insect cell medium can be included such as table 1 below institute
Show component.
The formula of an insect typical cell non-serum culture medium in the present invention of table 1
Present invention also offers application of the insect cell medium in culture insect cell or Prepare restructuring albumen.
Compared with prior art, the present invention at least has the following advantages that：The insect cell serum free medium component is simple
Clearly, easily prepared, cost is cheap, and Insect cellculture efficiency and expression of recombinant proteins efficiency can be substantially improved, beneficial to reality
The large-scale culture of existing insect cell and the extensive preparation of recombinant protein.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the required accompanying drawing used in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments described in invention, for those of ordinary skill in the art, on the premise of not paying creative work,
Other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 be with insect cell free serum culture in embodiment 1 based in 3L shaking flasks cultivate Sf9 cells culture density and
Survival curves figure；
Fig. 2 be with insect cell free serum culture in embodiment 1 based in 3L shaking flasks cultivate S2 cells culture density and
Survival curves figure；
Fig. 3 is based on cultivating Sf9 cells in 1L shaking flasks for respectively with insect cell free serum culture in embodiment 1, reference examples 1
Culture effect figure；
Fig. 4 is based on cultivating Sf9 cells in 1L shaking flasks for respectively with insect cell free serum culture in embodiment 1, reference examples 2
And carry out the electrophoresis photographs of exogenous protein expression；
Fig. 5 is bent with the culture density of insect cell serum free medium culture High5 cells in embodiment 1 and survival rate
Line chart；
Fig. 6 is with the culture density and survival rate of insect cell serum free medium culture bolworm cell in embodiment 1
Curve map；
Fig. 7 be with the culture density of insect cell serum free medium culture prodenia litura gonad cell in embodiment 1 and
Survival curves figure.
Embodiment
In view of deficiency of the prior art, inventor is able to propose the present invention's through studying for a long period of time and largely putting into practice
Technical scheme.Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out in detail
Description, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.Based on this hair
Embodiment in bright, the every other reality that those of ordinary skill in the art are obtained on the premise of creative work is not made
Example is applied, belongs to the scope of protection of the invention.
Present invention system provides the insect cell serum free medium of improvement a kind of, and it contains basic amino acid, inorganic
The materials such as salt, vitamin, protolysate, the growth of insect cell and the expression of recombinant protein can be supported.
Wherein, primary carbon source and energy of the carbohydrate such as glucose, the sucrose or maltose system as insect cell growth
Source.Contained carbohydrate content is advantageous to ooze during the fermentation in 6042~14222mg/L in the culture medium of the present invention
Voltage-controlled system is in 380~410mOsm/Kg thoroughly, so as to meet the growth of insect cell.
Wherein, amino acid is also that insect cell growth is necessary, in general, has 14 kinds of amino acid to be carried by culture medium
For.In the present invention, by by the amino acid concentration control in culture medium in 11200~29170mg/L, than existing no blood
Clear Insect culture medium, amino acid dosage reduce, but can reach quite or even more preferably culture effect substantially.
Wherein, vitamin is to promoting insect cell survival and propagation and cell attachment to have positive role.In the present invention,
By the way that the vitamine concentration in culture medium is controlled in 11.354~820.4mg/L.Wherein Choline Chloride and inositol are given birth to cell
Lipid material metabolism synthesis in growth process has a major impact, and can promote cell growth, and maintain cell in shape.
Other vitamins participate in cell metabolism as coenzyme or prothetic group mostly.It is metabolized in insect cell intracellular and is moved different from other lactations
The intracellular metabolism of thing cell, insect cell glycometabolism need substantial amounts of vitamin to join based on tricarboxylic acid cycle, in metabolic process
With, and the more general mammalian cell of insect cell propagation is fast, so as to increase the demand to vitamin.Basis in the present invention
Enzyme material in insect cell is quantified and integrates the research of small-molecule substance group vitamin is quantified, achieved preferably
Cell culture effect.
Wherein, an important function system of inorganic salts provides the osmotic pressure beneficial to insect cell growth, also helps simultaneously
Promote cell attachment and increase yield, and promote the infection of virus in some cases and replicate big.In the present invention, be by
The concentration of inorganic salts in culture medium is controlled in 5500.71~9510.33mg/L, and by barium chloride, lithium chloride, chlorination
Nickel controls in the range of 0.001~0.1mg/L, 0.005~0.02mg/L, 0.05~7mg/L respectively, wherein the nothing widely applied
Machine salt helps to maintain ionic equilibrium, helps cell to be used for matter transportation for this normal metabolism and ion channel.Wherein add
Although barium chloride, lithium chloride and nickel chloride be for promoting cell growth, this crime in many serum free mediums
A person of good sense is also found surprisingly that very much, when adding a small amount of barium chloride, lithium chloride, chlorine in the serum-free insect culture medium in the present invention
After changing nickel, its growth to cell does not have obvious facilitation, but can increase considerably inclusion disease in insect cell
Poison and free virus, and then the expression of foreign protein can be significantly increased.Preferably, these materials are in serum-free insect culture medium
In dosage can be：0.001~0.1mg/L of barium chloride, 0.005~0.02mg/L of lithium chloride, nickel chloride 0.05~
7mg/L。
Wherein, the effect of protolysate is the growth for promoting cell under absence of serum.The egg added in the present invention
White hydrolysate source is plant, the hydrolysate in other sources is instead of with material protolysate, especially yeast hydrolyate can
To substantially reduce the endotoxin content in culture medium.In the present invention, by by the concentration control of the protolysate in culture medium
System is in 3000mg/L~11000mg/L, completely can be with the growth of sertoli cell, and within the range in the concentration range
The expression and baculovirus high titer amplification of recombinant exogenous protein can be supported.And make mostly in existing serum free medium
With yeast hydrolyate, and function of cell growth can be promoted just with it, not only consider cell growth in the present invention, and will
Foreign recombinant proteins are expressed and leading indicator of the baculovirus titers as Screening of Media.
Certainly, Serum-free complete medium of the invention can also include other compositions, such as water as solvent etc..
And need to stress, inventor has found in studying for a long period of time and largely putting into practice, when in serum-free elder brother
Appropriate phosphoric acid ester material is added in worm culture medium, particularly adds a small amount of dilinoleoylphosphatidylcholine and distearyl
After phosphatidyl choline, expressing quantity can be significantly improved, this is very amazing.Preferably, two kinds of materials are in nothing
Dosage in serum Insect culture medium can be：0.5~15mg/L of dilinoleoylphosphatidylcholine, DSPC
0.1~10mg/L.
The serum-free insect culture medium ingredient species of the present invention is few, and content is low, prepares simply, cost is cheap, and energy
Enough reach and be commercialized the same effect of culture medium, there is good Commercial Prospect.
The serum-free insect culture medium of the present invention can be made by any suitable method known in the art.For example, can be with
Each composition for forming the culture medium is first dissolved in water with respective suitable concn, remixes and is filtered to remove nuisance therein
Matter, for example, some insoluble particulate matters etc. and be made aseptic culture medium.
Postscript, when using the medium culture cell of the present invention, it can also use any appropraite condition known in the art, example
Depending on such as condition about the same with conventional medium, the species that it can be according to the cell system to be cultivated etc..
Technical scheme is described in more detail below in conjunction with specific embodiment and accompanying drawing.
The formula of the insect cell serum free medium of embodiment 1 is as shown in table 2.
A kind of formula of insect cell serum free medium in the embodiment 1 of table 2
And the preparation process of the culture medium of the present embodiment can include：
First, raw material is weighed or measured according to table 3；
Secondly, concentrate is configured, including：
Prepare 1000 × concentrate 1：
By ammonium molybdate tetrahydrate 8mg, CoCL2 6H2O 10mg, copper chloride dihydrate 75mg, ferrous sulfate heptahydrate 50mg, four water chlorine
Change manganese 8mg, zinc chloride 10mg is dissolved in 100ml water.
Prepare 1000 × concentrate 2：
Biotin 10mg, D-VB5 calcium 20mg, VB12 cyanocobalamin 20mg, VB9 folic acid 20mg, i- inositol 50mg are dissolved in
In 100ml water.
Prepare 1000 × concentrate 3：
Barium chloride 5mg, lithium chloride 1mg are dissolved in 100ml water.
Finally, take the other materials listed by foregoing concentrate and table 3 to weigh in 10L containers, prepare 10L Liquid Cultures
Base.Its process can be：9.5L water is added into the container, stirring 30min dissolvings, the concentrate 1 of above-mentioned preparation is added, concentrates
Liquid 2, concentrate 3, each 10ml, stirring, pH value is adjusted to 6.2 using 10M NaOH solutions, is settled to 10L, filtration sterilization, lucifuge
Refrigerate stand-by.
The dosage of insect cell serum free medium key component in the embodiment 1 of table 3
Reference examples 1：In addition to dilinoleoylphosphatidylcholine, DSPC is not added, according to implementation
The essentially identical component of example 1 and mode configure serum free medium.
Reference examples 2：In addition to barium chloride, lithium chloride, nickel chloride is not added, according to component substantially the same manner as Example 1
Serum free medium is configured with mode.
Example 1, the culture medium of reference examples 1 insert in 1L shaking flasks the culture Sf9 cells that suspend, its culture effect point respectively
Not as shown in Figure 1, Figure 3.Although it can be found that dilinoleoylphosphatidylcholine, DSPC in culture medium most
Maxicell stand density influences not notable but maintains have significant impact to cytoactive.When lacking two sub- oleoylphosphatidyl courages
When alkali, DSPC, although cell maximum cell density reaches maximum thin without significant changes in cell
Cytoactive can reduce rapidly after born of the same parents' density.
It can be found that using the medium culture Sf9 cells of the embodiment 1, maximum viable cell density more than 1.3 ×
107Cells/ml, even better than commercialization culture medium suitable with commercialization medium culture.
Example 1, the culture medium of reference examples 2 insert in 1L shaking flasks the culture Sf9 cells that suspend respectively, and in cell density
Reach 1.5 × 106Virus inoculation (recombinant virus, expressing corresponding foreign protein) during cells/ml, 72h is cultivated, takes cell suspension
Western-blot detections are carried out, for its result as shown in figure 4, wherein No. 1 swimming lane is the culture medium of comparative example 2, No. 2 swimming lanes are control
The culture medium of example 1, No. 3 swimming lanes are the culture medium of embodiment 1, and M is MarKer (label).It can be seen that addition barium chloride,
Lithium chloride, nickel chloride can greatly improve exogenous protein expression amount.
In addition, inventor has also carried out serum-free using the culture medium of the embodiment 1 to remaining various insects cell
Experiments On Suspension Culture, including S2, High5, Sf21 cell line etc., its typical result of the test is as shown in Fig. 2, Fig. 5.
In addition, inventor also found, the culture medium of the embodiment 1 can also support the elder brother that a variety of country voluntarily establish
The serum free suspension culture of worm cell line, such as the bolworm cell system (referring to Fig. 6) that Fudan University in Shanghai is established, University Of Suzhou builds
Vertical prodenia litura ovary cell line (referring to Fig. 7).
Embodiment 2：Cell culture：Be inoculated with 3 liters of shaking flasks 1000 milliliters of cell suspensions (select the culture medium of embodiment 1,
Wherein contain cell 0.5 × 106Cells/ml), it is placed on the shaking table in biochemical cultivation case, 27 DEG C, 100 revs/min of rotating speed,
When cell density reaches 5 × 106Cells/ml, by its diluted passage, its real density of cell is 0.5 × 106Cells/ml.By 3
Rise density and reach 5 × 106The cell suspension of cells/ml is transferred in 42L bioreactors, then adds fresh embodiment
1 27 liters of culture medium, speed of agitator are 50 revs/min, and 27 DEG C of temperature, dissolved oxygen control 40%, pH value is controlled 6.2.When cell is close
Degree reaches 5 × 106During cells/ml, transfer them in 1000 liters of bioreactors, and add fresh embodiment 1 and cultivate
Base controls 20 revs/min of speed of agitator, 27 DEG C of temperature, dissolved oxygen 40%, pH value is 6.2, when cell density reaches to 300 liters
3.75×106Cells/ml, the fresh culture medium of embodiment 1 is added to 750 liters, while added according to millesimal ratio
Recombinant virus, control environmental condition is constant, cultivates 72 hours, and foreign recombinant proteins expression quantity, recombinant protein table are detected with ELISA
It is 150 mg/litres up to amount, realizes insect cell serum free medium high level expression foreign protein.
It should be noted that herein, term " comprising ", "comprising" or its any other variant are intended to non-row
His property includes, so that process, method, article or equipment including a series of elements not only include those key elements, and
And also include the other element being not expressly set out, or also include for this process, method, article or equipment institute inherently
Key element.In the absence of more restrictions, the key element limited by sentence "including a ...", it is not excluded that including institute
State in process, method, article or the equipment of key element and other identical element also be present.
Described above is only the embodiment of the present invention, it is clear that described embodiment is only the present invention one
Divide embodiment, rather than whole embodiments.It should be pointed out that for those skilled in the art, do not taking off
On the premise of from the principle of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as the present invention's
Protection domain.
Claims (1)
1. a kind of insect cell free serum culture is based on the application in culture insect cell, it is characterised in that the insect cell
Serum free medium is made up of following component：
Beta-alanine 500mg/L
L- R-genes 900mg/L
Altheine 1000mg/L
L-Aspartic acid sodium salt 1000mg/L
CYSTINE hydrochloride 200mg/L
Pidolidone hydrochloride 1500mg/L
Glu 1000mg/L
Glycine 400mg/L
L-Histidine 500mg/L
L- hydroxyprolines 500mg/L
ILE 500mg/L
L-Leu 300mg/L
LYS 600mg/L
L-Methionine 800mg/L
L-phenylalanine 900mg/L
L-PROLINE 600mg/L
Serine 500mg/L
L-threonine 700mg/L
L-Trp 300mg/L
TYR disodium salt 750mg/L
Valine 800mg/L
Ammonium molybdate tetrahydrate 0.08mg/L
CoCL2 6H2O 0.1mg/L
Copper chloride dihydrate 0.75mg/L
Ferrous sulfate heptahydrate 0.5mg/L
Anhydrous magnesium sulfate 900mg/L
Tetrahydrate manganese chloride 0.08mg/L
Potassium chloride 1500mg/L
Sodium chloride 4000mg/L
Sodium dihydrogen phosphate-water 1000mg/L
Zinc chloride 0.1mg/L
Biotin 0.1mg/L
D-VB5 calcium 0.2mg/L
Choline Chloride 10mg/L
VB12 cyanocobalamins 0.2mg/L
VB9 folic acid 0.2mg/L
I- inositols 0.5mg/L
VB3 nicotinic acid 1mg/L
P-aminobenzoic acid 2mg/L
VB6 puridoxine hydrochlorides 1mg/L
VB2 Riboflavin Tetrabutyrates mg/L
VB1 thiamine hydrochlorides 2mg/L
Barley hydrolysate 4000mg/L
Rice hydrolysate 4000mg/L
DEXTROSE ANHYDROUS 5000mg/L
Fumaric acid 5mg/L
2-oxoglutaric acid 20mg/L
L MALIC ACID 20mg/L
Ultrapure water Fructus Hordei Germinatus sugar 500mg/L
Succinic acid 4mg/L
Tween80 100mg/L
Cholesterol sulfate ester sodium salt 15mg/L
VE sodium 2.5mg/L
D- sucrose 3000mg/L
Dilinoleoylphosphatidylcholine 2mg/L
DSPC 1.25mg/L
Barium chloride 0.05mg/L
Lithium chloride 0.01mg/L
Nickel chloride 3.5mg/L.
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| CN105255811A (en) * | 2015-11-09 | 2016-01-20 | 内蒙古金源康生物工程有限公司 | Preparing method for serum-free and animal-source-free culture medium additive suitable for growth of insect cells |
| CN105255810A (en) * | 2015-11-09 | 2016-01-20 | 内蒙古金源康生物工程有限公司 | Serum-free and animal-source-free culture medium suitable for insect cell Sf-9 |
| SG11201806335RA (en) * | 2016-02-22 | 2018-09-27 | Agency Science Tech & Res | Cell culture medium |
| CN105861416B (en) * | 2016-06-06 | 2019-08-02 | 西北民族大学 | It is a kind of for the serum free medium and its preparation method and application for cultivating insect cell that suspends entirely |
| CN106367379B (en) * | 2016-12-07 | 2019-09-10 | 天康生物股份有限公司 | A kind of insect cell maintaining liquid and its preparation method improving swine fever virus E2 recombinant protein expression quantity |
| CN106834203A (en) * | 2016-12-24 | 2017-06-13 | 严志海 | A kind of insect cell medium |
| CN110564670A (en) * | 2019-09-04 | 2019-12-13 | 广州今成生物科技有限公司 | Insect cell serum-free culture medium and preparation process thereof |
| CN114045258B (en) * | 2021-10-21 | 2024-04-16 | 辽宁盛京干细胞科技有限公司 | Serum-free culture medium for mesenchymal stem cell culture and application |
| CN113832094B (en) * | 2021-10-29 | 2024-04-16 | 无锡多宁生物科技有限公司 | Basal medium without serum and animal source components of insect cells and preparation method thereof |
| CN115627250B (en) * | 2022-12-01 | 2023-04-04 | 天信和(苏州)生物科技有限公司 | Serum-free medium for culturing insect cells and application thereof |
| CN116478903B (en) * | 2023-05-08 | 2023-10-24 | 苏州依科赛生物科技股份有限公司 | Insect cell serum-free culture medium and application thereof |
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| CN101988047A (en) * | 2010-05-13 | 2011-03-23 | 维亚生物科技(上海)有限公司 | Insect cell serum-free medium with low cost |
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