CN115627250B - Serum-free medium for culturing insect cells and application thereof - Google Patents

Serum-free medium for culturing insect cells and application thereof Download PDF

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CN115627250B
CN115627250B CN202211523747.6A CN202211523747A CN115627250B CN 115627250 B CN115627250 B CN 115627250B CN 202211523747 A CN202211523747 A CN 202211523747A CN 115627250 B CN115627250 B CN 115627250B
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concentration
virus
serum
acid
vitamin
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CN115627250A (en
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陈文庆
徐舸辰
赵洪磊
浦勇
类成霞
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Tianxinhe Suzhou Biotechnology Co ltd
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Abstract

The invention provides a serum-free culture medium for culturing insect cells and application thereof, wherein the serum-free culture medium for culturing the insect cells does not contain serum components, the insect cells are cultured by adopting the serum-free culture medium, high-density culture of the insect cells is supported, and the insect cells or viruses are amplified and cultured by adopting the serum-free culture medium, so that the obtained product can reduce the risk of initiating the immune reaction of an animal body, has the advantages of high safety and the like, is convenient for the separation and purification of subsequent products, and is more suitable for industrial production.

Description

Serum-free medium for culturing insect cells and application thereof
Technical Field
The invention relates to the technical field of biology, in particular to a serum-free culture medium for culturing insect cells and application thereof, and more particularly relates to a serum-free culture medium for culturing insect cells, a method for amplifying viruses, a vaccine and application thereof.
Background
Insect cells are eukaryotic cells from insects, can express proteins at high level by culturing the insect cells, and gene segments of the insect cells can be used for constructing recombinant viruses to efficiently express foreign genes and can be used for producing virus vaccines. Moreover, the first recombinant protein corona vaccine produced by insect cells has entered phase two clinical trials. Therefore, the in vitro culture of insect cells is increasingly receiving attention.
However, at present, a culture medium containing serum is often used for amplification culture of insect cells, but the application of serum has many problems, for example, serum may cause an immune response in an animal body, and the presence of a large amount of serum proteins increases the difficulty of vaccine separation and purification. Therefore, development of a serum-free medium for culturing insect cells has been continued.
Disclosure of Invention
The present invention aims to address at least some of the above technical problems or at least to provide a useful commercial choice. Therefore, the invention provides a serum-free medium capable of effectively culturing insect cells.
Thus, in a first aspect of the invention, the invention provides a serum-free medium for culturing insect cells. According to an embodiment of the invention, the serum-free medium comprises: IPL41 medium and additives; the additive comprises the following components: 2700 parts by weight of pea hydrolysate, 0.26352 parts by weight of copper sulfate pentahydrate, 0.495 parts by weight of ferrous sulfate heptahydrate, 4050 parts by weight of D-anhydrous glucose, 777.6 parts by weight of glutamine dipeptide, 0.9 parts by weight of soybean lecithin, 900 parts by weight of F68,4.32 parts by weight of succinic acid, 0.144 parts by weight of p-aminobenzoic acid, 0.036 parts by weight of D-biotin, 0.4977 parts by weight of calcium D-pantothenate, 10.197 parts by weight of choline chloride, 0.6507 parts by weight of folic acid, 6.1137 parts by weight of inositol, 0.1827 parts by weight of vitamin B6,0.126 parts by weight of vitamin B2,0.5571 parts by weight of vitamin B1,1.1889 parts by weight of vitamin B12, 33.03 parts by weight of sodium pyruvate, 0.0999 parts by weight of glutathione, 710.55 parts by weight of L-asparagine, 234.81 parts by weight of L-cysteine hydrochloride, 10.224 parts by weight of L-cystine hydrochloride, 123.3 parts by weight of L-serine, 1.71 parts by weight of L-threonine, 60.3 parts by weight of L-valine, 4500 parts by weight of yeast hydrolysate, 2700 parts by weight of soybean hydrolysate, 0.603 parts by weight of vitamin C,0.5202 parts by weight of nicotinamide, 0.0261 parts by weight of pyridoxal hydrochloride, 16.2 parts by weight of magnesium chloride hexahydrate, 0.0001 parts by weight of ethanolamine, 0.0012 parts by weight of hydrocortisone, 0.0295 parts by weight of lipoic acid, 0.004 parts by weight of sodium selenite, 0.469 parts by weight of 1, 4-butanediamine dihydrochloride, 2.23E-06 parts by weight of T3,9.05E-07 parts by weight of sodium metavanadate, 9.71E-06 parts by weight of germanium dioxide, 1.23E-05 parts by weight of potassium bromide, 0.0001 parts by weight of ammonium metavanadate, 0.0001 parts by weight of rubidium chloride, 0.0002 parts by weight of cadmium chloride, 0.0003.0003 parts by weight of barium chloride, 3.0003 parts by weight of barium chloride, 0.0004 part by weight of sodium fluoride, 0.001 part by weight of copper chloride dihydrate, 0.0021 part by weight of lithium chloride, 0.021 part by weight of aluminum chloride hexahydrate, 0.078 part by weight of sodium metasilicate nonahydrate, 0.2097 part by weight of hypoxanthine, 0.04 part by weight of thymidine, 0.36 part by weight of insulin, 3.98 parts by weight of ferric citrate, 1.98 parts by weight of cholesterol, 0.63 part by weight of vitamin E acetate, 0.09 part by weight of linoleic acid, 0.09 part by weight of myristic acid, 0.09 part by weight of oleic acid, 0.09 part by weight of palmitic acid, 0.09 part by weight of stearic acid, and 19.8 parts by weight of tween 80.
According to the embodiment of the invention, the additive for culturing insect cells is optimized and adjusted through a large number of experiments to obtain the culture medium without serum, and the inventor does not find the rule among the components in the additive in the experiment process, and the result is unpredictable. However, the inventor finds in experiments that the high-density culture of insect cells is supported by adopting the serum-free culture medium to culture the insect cells, and the survival rate of the cells can reach more than 99 percent; compared with the method for amplifying and culturing insect cells or viruses by adopting a culture medium containing serum, particularly the amplification production of virus vaccines, the obtained product can reduce the risk of triggering the immune reaction of animal organisms, has the advantages of high safety and the like, is convenient for the separation and purification of subsequent products, and is more suitable for industrial production. The serum-free culture medium containing the additive has the advantages of small difference between batches, strong repeatability, stable result, no animal source risk and the like.
According to the embodiment of the invention, based on the total volume of the serum-free culture medium, the concentration of pea hydrolysate is 2700mg/L, the concentration of copper sulfate pentahydrate is 0.26352mg/L, the concentration of ferrous sulfate heptahydrate is 0.495mg/L, the concentration of D-anhydrous glucose is 4050mg/L, the concentration of glutamine dipeptide is 777.6mg/L, the concentration of soybean lecithin is 0.9mg/L, the concentration of F68 is 900mg/L, the concentration of succinic acid is 4.32mg/L, the concentration of p-aminobenzoic acid is 0.144mg/L, the concentration of D-biotin is 0.036mg/L, the concentration of D-calcium pantothenate is 0.4977mg/L, the concentration of choline chloride is 10.197mg/L, the concentration of folic acid is 0.6507mg/L, the concentration of inositol is 6.1137mg/L, the concentration of vitamin B6 is 0.1827mg/L, and the concentration of vitamin B2 is 0.126mg/L, the concentration of vitamin B1 is 0.5571mg/L, the concentration of vitamin B12 is 1.1889mg/L, the concentration of sodium pyruvate is 33.03mg/L, the concentration of glutathione is 0.0999mg/L, the concentration of L-asparagine is 710.55mg/L, the concentration of L-cysteine hydrochloride monohydrate is 234.81mg/L, the concentration of L-cystine dihydrochloride is 10.224mg/L, the concentration of L-serine is 123.3mg/L, the concentration of L-threonine is 1.71mg/L, the concentration of L-valine is 60.3mg/L, the concentration of yeast hydrolysate is 4500mg/L, the concentration of soybean hydrolysate is 2700mg/L, the concentration of vitamin C is 0.603mg/L, the concentration of nicotinamide is 0.5202mg/L, the concentration of pyridoxal hydrochloride is 0.0261mg/L, and the concentration of magnesium chloride hexahydrate is 16.2mg/L, the concentration of ethanolamine is 0.0001mg/L, the concentration of hydrocortisone is 0.0012mg/L, the concentration of lipoic acid is 0.0295mg/L, the concentration of sodium selenite is 0.004mg/L, the concentration of 1, 4-butanediamine dihydrochloride is 0.469mg/L, the concentration of T3 is 2.23E-06mg/L, the concentration of sodium metavanadate is 9.05E-07mg/L, the concentration of germanium dioxide is 9.71E-06mg/L, the concentration of potassium bromide is 1.23E-05mg/L, the concentration of ammonium metavanadate is 0.0001mg/L, the concentration of rubidium chloride is 0.0001mg/L, the concentration of cadmium chloride is 0.0002mg/L, the concentration of barium acetate is 0.0003mg/L, the concentration of zirconium octoate is 0.0003mg/L, the concentration of sodium fluoride is 0.4 mg/L, the concentration of copper chloride dihydrate is 0.001mg/L, the concentration of lithium chloride is 0.0021mg/L, the concentration of aluminum chloride hexahydrate is 0.021mg/L, the concentration of sodium metasilicate nonahydrate is 0.078mg/L, the concentration of hypoxanthine is 0.2097mg/L, the concentration of thymidine is 0.04mg/L, the concentration of insulin is 0.36mg/L, the concentration of ferric citrate is 3.98mg/L, the concentration of cholesterol is 1.98mg/L, the concentration of vitamin E acetate is 0.63mg/L, the concentration of linoleic acid is 0.09mg/L, the concentration of myristic acid is 0.09mg/L, the concentration of oleic acid is 0.09mg/L, the concentration of palmitic acid is 0.09mg/L, the concentration of stearic acid is 0.09mg/L, and the concentration of Tween 80 is 19.8mg/L. Thereby, the cell density and the cell survival rate of the insect cells can be further improved.
According to an embodiment of the invention, the IPL41 medium comprises the following components:
Figure 744804DEST_PATH_IMAGE001
Figure 337591DEST_PATH_IMAGE002
according to an embodiment of the invention, the insect cells comprise at least one selected from the group consisting of SF9 cells, SF21 cells and High Five cells.
In a second aspect of the invention, a method of expanding insect cells is provided. According to an embodiment of the invention, the method comprises: culturing said insect cells in a serum-free medium according to the first aspect, such that expansion of said insect cells is achieved. According to the embodiment of the invention, the method can support high-density suspension culture of insect cells, and the survival rate of the insect cells can reach more than 99%.
In a third aspect of the invention, a method of amplifying a virus is provided. According to an embodiment of the invention, the method comprises: performing amplification culture on insect cells by using the method of the second aspect to obtain a culture solution; inoculating the culture with a virus to effect amplification of the virus. According to the embodiment of the present invention, the virus can be efficiently amplified by the above method.
According to an embodiment of the present invention, the virus includes at least one selected from the group consisting of rabies virus, influenza virus, coronavirus, bursa of fabricius virus, dengue virus, enterovirus, encephalitis virus, poxvirus, orthomyxovirus, paramyxovirus, retrovirus, togavirus, flavivirus, enterovirus, picornavirus, mosaic virus, herpes virus, adenovirus, vaccinia virus, SARS virus, influenza a virus, influenza b virus, lentivirus, ross river virus, west nile virus, yellow fever virus, FSME virus, and hepatitis a virus.
Illustratively, the vaccines include, but are not limited to, swine fever vaccine, porcine diarrhea vaccine, porcine pseudorabies vaccine, and neocorona vaccine.
In a fourth aspect of the invention, a vaccine is presented. According to an embodiment of the invention, the vaccine is obtained by attenuating or inactivating the virus according to the third aspect. According to the embodiment of the invention, the vaccine does not contain serum, can reduce the risk of inducing immune response of animal bodies, and has the advantages of high safety and the like.
In a fifth aspect, the present invention provides the use of a vaccine according to the fourth aspect in the manufacture of a medicament for the treatment or prophylaxis of a virus-related disease.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 shows the results of cell density of insect cells cultured in different media for 72 hours according to example 1 of the present invention;
FIG. 2 shows the results of the cell viability of insect cells cultured in different media for 72 hours in example 1 of the present invention;
FIG. 3 is a graph showing the expression level of porcine circovirus capsid protein after inoculation of porcine circovirus with insect cells cultured in different media according to example 1 of the present invention, wherein lane 1 shows the results for serum-free medium B, lane 2 shows the results for serum-free medium A, lane 4 shows the results for serum-free medium, and lane 7 shows the results for commercial medium.
Detailed Description
The following detailed description of embodiments of the invention is intended to be illustrative, and not to be construed as limiting the invention.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In this document, the terms "comprise" or "comprise" are open-ended expressions that include the elements indicated in the present invention, but do not exclude other elements.
In this context, the term "treatment" is intended to mean the use to obtain a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of complete or partial prevention of the disease or symptoms thereof, and/or may be therapeutic in terms of a partial or complete cure for the disease and/or adverse effects resulting from the disease. As used herein, "treatment" encompasses diseases in mammals, particularly humans, including: (a) Preventing the occurrence of a disease or disorder in an individual susceptible to the disease but not yet diagnosed; (b) inhibiting a disease, e.g., arresting disease progression; or (c) alleviating the disease, e.g., alleviating symptoms associated with the disease. As used herein, "treatment" encompasses any administration of a drug or compound to an individual to treat, cure, alleviate, ameliorate, reduce or inhibit a disease in the individual, including but not limited to the administration of a drug containing a compound described herein to an individual in need thereof.
The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or apparatus used are conventional products which are commercially available, not indicated by the manufacturer, and the following materials are used in the examples: pea hydrolysate (chikayabe, cat # a 2501), soy hydrolysate (chikayabe, a 1603), yeast hydrolysate (chikayabe, a 1202).
Example 1
1. Preparation of culture Medium
Serum-free medium a (a serum-free medium to which the present invention was applied) and serum-free medium B (a control serum-free medium) were obtained by adding an additive to IPL41 medium (commercially available, see table 1 for a specific formulation) at the concentrations of the respective components of the additive in serum-free medium a or serum-free medium B, respectively, according to table 2. A medium containing IPL41+10% NBS (newborn bovine serum) was prepared as a serum medium, and a commercially available medium (Volmer H502C) was prepared as a control.
Table 1: IPL41 culture medium formula
Name of raw materials mg/L
Glycine 200
L-hydroxyproline 800
L-arginine hydrochloride 800
L-asparagine 1300
L-aspartic acid 1300
L-cystine dihydrochloride 119.14
L-glutamic acid 1500
L-Glutamine 1000
L-histidine hydrochloride monohydrate 200
L-isoleucine 750
L-leucine 250
L-lysine hydrochloride 700
L-methionine 1000
L-phenylalanine 1000
L-proline 500
L-serine 200
L-threonine 200
L-tryptophan 100
L-tyrosine disodium salt 360.4
L-valine 500
Beta-alanine 300
D-biotin 0.16
Choline chloride 20
D-calcium pantothenate 0.008
Folic acid 0.08
Nicotinic acid 0.16
Para aminobenzoic acid 0.32
Vitamin B6 0.4
Vitamin B2 0.08
Succinic acid 4.8
Vitamin B1 0.08
Vitamin B12 0.24
Inositol 0.4
Ammonium molybdate tetrahydrate 0.04
Calcium chloride dihydrate 500
Cobalt chloride hexahydrate 0.05
Cupric chloride dihydrate 0.2
Iron sulfate heptahydrate 0.55
Anhydrous magnesium sulfate 918
Manganese chloride tetrahydrate 0.02
Potassium chloride 1200
Sodium bicarbonate 350
Sodium chloride 2850
Monobasic sodium phosphate monohydrate 1160
Zinc chloride 0.04
Alpha-ketoglutaric acid 29.6
D-Anhydrous glucose 2500
Fumaric acid 4.4
L-malic acid 53.6
Maltose 1000
Sucrose 1650
Table 2: formulation and addition amount of each additive in serum-free medium A and serum-free medium B
Raw material name of additive Serum-free medium A (mg/L) Serum-free medium B (mg/L)
Pea hydrolysate 2700 2000
Blue vitriod 0.26352 0.26352
Ferrous sulfate heptahydrate 0.495 0.495
D-Anhydrous glucose 4050 3000
Glutamine dipeptide 777.6 458.78
Soybean lecithin 0.9 0.5
F68 900 600
Succinic acid 4.32 3.5
Para aminobenzoic acid 0.144 0.0864
D-biotin 0.036 0.0216
D-calcium pantothenate 0.4977 0.29862
Choline chloride 10.197 6.1182
Folic acid 0.6507 0.39042
Inositol 6.1137 3.66822
Vitamin B6 0.1827 0.14616
Vitamin B2 0.126 0.1008
Vitamin B1 0.5571 0.44568
Vitamin B12 1.1889 0.95112
Pyruvic acid sodium salt 33.03 19.818
Glutathione 0.0999 0.07992
L-asparagine 710.55 348.1695
L-cysteine hydrochloride monohydrate 234.81 115.0569
L-cystine dihydrochloride 10.224 5.00976
L-serine 123.3 60.417
L-threonine 1.71 0.8379
L-valine 60.3 29.547
Yeast hydrolysate 4500 3000
Soybean hydrolysate 2700 3000
Vitamin C 0.603 0.4824
Nicotinamide 0.5202 0.41616
Pyridoxal hydrochloride 0.0261 0.02088
Magnesium chloride hexahydrate 16.2 15
Ethanolamine 0.0001 7.44972E-05
Hydrocortisone 0.0012 0.000983363
Lipoic acid 0.0295 0.02359657
Sodium selenite 0.0040 0.003194028
1, 4-butanediamine dihydrochloride 0.4690 0.229826673
T3 2.23E-06 1.78793E-06
Sodium metavanadate 9.05E-07 7.24233E-07
Germanium dioxide 9.71131E-06 7.76905E-06
Potassium bromide 1.23324E-05 9.86593E-06
Ammonium metavanadate 0.0001 9.86952E-05
Rubidium chloride 0.0001 0.000100186
Cadmium chloride 0.0002 0.000151578
Barium acetate 0.0003 0.000211237
Zirconium oxychloride octahydrate 0.0003 0.000266895
Sodium fluoride 0.0004 0.000348017
Cupric chloride dihydrate 0.0010 0.00076242
Lithium chloride 0.0021 0.001645984
Aluminium chloride hexahydrate 0.0210 0.016808619
Sodium metasilicate nonahydrate 0.0780 0.062375688
Hypoxanthine 0.2097 0.16776
Thymidine 0.04 0.03120392
Insulin 0.36 0.284780921
Ferric citrate 3.98 2.385040212
Cholesterol 1.98 1.188
Vitamin E acetate 0.63 0.378
Linoleic acid 0.09 0.054
Myristic acid 0.09 0.054
Oleic acid 0.09 0.054
Palmitic acid 0.09 0.054
Stearic acid 0.09 0.054
Tween 80 19.8 11.88
2. Cell culture and detection
After serum-free medium A, serum-free medium B, serum medium and commercial medium were obtained as described above, insect cells (SF 9, purchased from ATCC, cat # CRL-1711) were cultured in the following manner at an initial cell density of 0.7X 10 6 cells/ml, culture conditions: 27 ℃ C., 120rpm, 250ml shake flask (vented lid). After 72h of culture, the cell density and cell viability of the insect cells were examined and serial passages were performed. After adapting to 5 generations, the cells were examined for growth stability by continuous passage, and the cell density and cell viability are shown in FIGS. 1 and 2. The cell density and the cell survival rate of the serum-free culture medium are superior to those of the serum culture medium and superior to commercial culture media.
3. Viral infection and detection
And inoculating porcine circovirus (PCV 2) to the insect cells growing in the logarithmic phase cultured in the different culture media according to MOI 0.05, and detecting the PCV2 capsid protein expression quantity in the insect cells by Western Blot 48h after inoculation. As shown in FIG. 3, the protein expression of serum-free culture A was comparable to that of commercial culture medium, and both of them were superior to that of serum culture medium.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made in the above embodiments by those of ordinary skill in the art without departing from the principle and spirit of the present invention.

Claims (5)

1. A serum-free medium for culturing insect cell SF9 cells, comprising:
IPL41 medium and additives;
the additive consists of the following components:
based on the total volume of the serum-free culture medium, the concentration of pea hydrolysate is 2700mg/L, the concentration of blue vitriol is 0.26352mg/L, the concentration of ferrous sulfate heptahydrate is 0.495mg/L, the concentration of D-anhydrous glucose is 4050mg/L, the concentration of glutamine dipeptide is 777.6mg/L, the concentration of soybean lecithin is 0.9mg/L, the concentration of F68 is 900mg/L, the concentration of succinic acid is 4.32mg/L, the concentration of p-aminobenzoic acid is 0.144mg/L, the concentration of D-biotin is 0.036mg/L, the concentration of D-calcium pantothenate is 0.4977mg/L, and the concentration of choline chloride is 10.197mg/L, the concentration of folic acid is 0.6507mg/L, the concentration of inositol is 6.1137mg/L, the concentration of vitamin B6 is 0.1827mg/L, the concentration of vitamin B2 is 0.126mg/L, the concentration of vitamin B1 is 0.5571mg/L, the concentration of vitamin B12 is 1.1889mg/L, the concentration of sodium pyruvate is 33.03mg/L, the concentration of glutathione is 0.0999mg/L, the concentration of L-asparagine is 710.55mg/L, the concentration of L-cysteine hydrochloride monohydrate is 234.81mg/L, the concentration of L-cystine dihydrochloride is 10.224mg/L, the concentration of L-serine is 123.3mg/L, and the concentration of L-threonine is 1.71mg/LThe concentration of L-valine is 60.3mg/L, the concentration of yeast hydrolysate is 4500mg/L, the concentration of soybean hydrolysate is 2700mg/L, the concentration of vitamin C is 0.603mg/L, the concentration of nicotinamide is 0.5202mg/L, the concentration of pyridoxal hydrochloride is 0.0261mg/L, the concentration of magnesium chloride hexahydrate is 16.2mg/L, the concentration of ethanolamine is 0.0001mg/L, the concentration of hydrocortisone is 0.0012mg/L, the concentration of lipoic acid is 0.0295mg/L, the concentration of sodium selenite is 0.004mg/L, the concentration of 1, 4-butanediamine dihydrochloride is 0.469mg/L, and the concentration of T3 is 2.23X 10 -6 mg/L, concentration of sodium metavanadate is 9.05X 10 -7 mg/L, the concentration of germanium dioxide is 9.71X 10 -6 mg/L, concentration of potassium bromide 1.23X 10 -5 mg/L, the concentration of ammonium metavanadate is 0.0001mg/L, the concentration of rubidium chloride is 0.0001mg/L, the concentration of cadmium chloride is 0.0002mg/L, the concentration of barium acetate is 0.0003mg/L, the concentration of zirconium oxychloride octahydrate is 0.0003mg/L, the concentration of sodium fluoride is 0.0004mg/L, the concentration of copper chloride dihydrate is 0.001mg/L, the concentration of lithium chloride is 0.0021mg/L, the concentration of aluminum chloride hexahydrate is 0.021mg/L, the concentration of sodium metasilicate nonahydrate is 0.078mg/L, the concentration of hypoxanthine is 0.2097mg/L, the concentration of thymidine is 0.04mg/L, the concentration of insulin is 0.36mg/L, the concentration of ferric citrate is 3.98mg/L, the concentration of cholesterol is 1.98mg/L, the concentration of vitamin E acetate is 0.63mg/L, the concentration of linoleic acid is 0.09mg/L, the concentration of myristic acid is 0.09mg/L, the concentration of oleic acid is 0.09mg/L, the concentration of palmitic acid is 0.09mg/L, the concentration of stearic acid is 0.09mg/L, and the concentration of Tween 80 is 19.8mg/L.
2. A method of expanding an insect cell SF9 cell comprising:
culturing the insect cell SF9 cell in the serum-free medium of claim 1 to expand the insect cell.
3. A method of amplifying a virus, comprising:
performing amplification culture on insect cells by using the method of claim 2 to obtain a culture solution;
inoculating the culture with a virus to effect amplification of the virus.
4. The method of claim 3, wherein the virus comprises at least one selected from the group consisting of rabies virus, influenza virus, coronavirus, bursal virus, enterovirus, encephalitis virus, poxvirus, orthomyxovirus, paramyxovirus, retrovirus, togavirus, flavivirus, picornavirus, mosaic virus, herpesvirus, adenovirus, ross river virus, west Nile river virus, FSME virus, and hepatitis A virus.
5. The method of claim 3, wherein the virus comprises a virus selected from the group consisting of SARS virus, influenza A virus, influenza B virus, lentivirus, vaccinia virus, dengue virus, yellow fever virus.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101195816A (en) * 2007-12-28 2008-06-11 天津百若克医药生物技术有限责任公司 Gonad cell amplification culture medium of Chinese hamster and uses thereof
CN102925486A (en) * 2011-12-26 2013-02-13 武汉中博生物股份有限公司 Porcine circovirus type 2 subunit vaccine, and preparation method and application thereof
CN103122352A (en) * 2012-09-27 2013-05-29 华中农业大学 Porcine circovirus II-type recombinant baculovirus as well as preparation method and application thereof
CN104593316A (en) * 2015-02-02 2015-05-06 苏州市沃美生物技术有限公司 Insect cell serum-free culture medium and application thereof
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
CN112961825A (en) * 2021-02-26 2021-06-15 江苏普瑞康生物医药科技有限公司 Serum-free medium and preparation method thereof
CN115386537A (en) * 2022-10-28 2022-11-25 天信和(苏州)生物科技有限公司 Chemically defined media additives for Vero cells, and their use in culturing cells and amplifying viruses

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101195816A (en) * 2007-12-28 2008-06-11 天津百若克医药生物技术有限责任公司 Gonad cell amplification culture medium of Chinese hamster and uses thereof
CN102925486A (en) * 2011-12-26 2013-02-13 武汉中博生物股份有限公司 Porcine circovirus type 2 subunit vaccine, and preparation method and application thereof
CN103122352A (en) * 2012-09-27 2013-05-29 华中农业大学 Porcine circovirus II-type recombinant baculovirus as well as preparation method and application thereof
CN104593316A (en) * 2015-02-02 2015-05-06 苏州市沃美生物技术有限公司 Insect cell serum-free culture medium and application thereof
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
CN112961825A (en) * 2021-02-26 2021-06-15 江苏普瑞康生物医药科技有限公司 Serum-free medium and preparation method thereof
CN115386537A (en) * 2022-10-28 2022-11-25 天信和(苏州)生物科技有限公司 Chemically defined media additives for Vero cells, and their use in culturing cells and amplifying viruses

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