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Serum-free culture medium for culturing DF-1 cells and preparation method of serum-free culture medium

A serum-free medium, DF-1 technology, applied in the field of serum-free medium for culturing DF-1 cells and its preparation, can solve the problems of low cell concentration, low virus titer, cumbersome culture process, etc. Ease of use, improved product quality, high cell density

Pending Publication Date: 2021-01-12
山东信得动物疫苗有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the most effective preventive measures are vaccines. The key to preparing high-efficiency vaccines is to obtain high-titer viruses. Traditional vaccine production is to culture DF-1 cells on the wall with serum-containing medium and then inoculate them. The cell concentration is low, the virus titer is low, high concentration is required to meet the requirements of vaccine preparation, and the cultivation process is cumbersome, which cannot meet the needs of large-scale vaccine production
There is no commercially available serum-free medium for DF-1 cells, and studies on serum-free medium for DF-1 cells have not been reported

Method used

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  • Serum-free culture medium for culturing DF-1 cells and preparation method of serum-free culture medium
  • Serum-free culture medium for culturing DF-1 cells and preparation method of serum-free culture medium
  • Serum-free culture medium for culturing DF-1 cells and preparation method of serum-free culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] This specific example provides a serum-free medium for culturing DF-1 cells, which consists of amino acids, vitamins, salts, lipids, trace elements, buffers, protein hydrolyzates, acid-base indicators and other additives grouped into, among them,

[0050] Amino acids are alanine, arginine, asparagine, aspartic acid, cystine, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine , lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine;

[0051] The vitamins are biotin, folic acid, niacinamide, pyridoxine, thiamine and lipoic acid;

[0052] The salts are magnesium chloride, magnesium sulfate, calcium chloride, potassium chloride, sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium pyruvate;

[0053] The lipids are cholesterol, tocopheryl acetate, myristic acid, palmitic acid, palmitoleic acid, stearic acid, Tween and block polyether F68;

[0054] Trace elements are copper sul...

Embodiment 2

[0071] This specific example provides a serum-free medium for culturing DF-1 cells, and the specific content of each component in the serum-free medium for cultivating DF-1 cells is:

[0072] Amino acid part:

[0073] Alanine 4 mg / L,

[0074] Arginine 100 mg / L,

[0075] Asparagine 6 mg / L,

[0076] Aspartic Acid 9 mg / L,

[0077] Cystine 15 mg / L,

[0078] Cysteine ​​20mg / L,

[0079] Glutamic acid 7 mg / L,

[0080] Glutamine 200 mg / L,

[0081] Glycine 10 mg / L,

[0082] Histidine 20 mg / L,

[0083] Isoleucine 30 mg / L,

[0084] Leucine 50 mg / L,

[0085] Lysine 100mg / L,

[0086] Methionine 15 mg / L,

[0087] Phenylalanine 30 mg / L,

[0088] Proline 15 mg / L,

[0089] Serine 20 mg / L,

[0090] Threonine 30mg / L,

[0091] Tryptophan 15 mg / L,

[0092] Tyrosine 40mg / L,

[0093] Valine 40mg / L.

[0094] The content of described vitamin is:

[0095] Biotin 0.003 mg / L,

[0096] Folic acid 2 mg / L,

[0097] Niacinamide 5mg / L,

[0098] Pyridoxine 2 mg / L,

[0099] Thiamine 2 mg...

Embodiment 3

[0138] This specific example provides another serum-free medium for culturing DF-1 cells, and the specific content of each component in the serum-free medium for cultivating DF-1 cells is:

[0139] Amino acid part:

[0140] Alanine 8 mg / L,

[0141] Arginine 200mg / L,

[0142] Asparagine 20 mg / L,

[0143] Aspartic Acid 16 mg / L,

[0144] Cystine 25 mg / L,

[0145] Cysteine ​​35 mg / L,

[0146] Glutamic acid 15mg / L,

[0147] Glutamine 800mg / L,

[0148] Glycine 25 mg / L,

[0149] Histidine 50 mg / L,

[0150] Isoleucine 60mg / L,

[0151] Leucine 70mg / L,

[0152] Lysine 150 mg / L,

[0153] Methionine 30 mg / L,

[0154] Phenylalanine 50 mg / L,

[0155] Proline 30mg / L,

[0156] Serine 40mg / L,

[0157] Threonine 60mg / L,

[0158] Tryptophan 30 mg / L,

[0159] Tyrosine 70 mg / L,

[0160] Valine 70mg / L.

[0161] The content of described vitamin is:

[0162] Biotin 0.01 mg / L,

[0163] Folic acid 5 mg / L,

[0164] Niacinamide 10mg / L,

[0165] Pyridoxine 5 mg / L,

[0166] Thiamine...

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Abstract

The invention discloses a serum-free culture medium for culturing DF-1 cells and a preparation method of the serum-free culture medium. The serum-free culture medium for culturing DF-1 cells comprisesamino acids, vitamins, salts, lipids, trace elements, a buffer agent, protein hydrolysate, an acid-base indicator and other additive components, and the culture medium can be suitable for full-suspension serum-free culture of DF-1 cells by limiting specific concentrations of various components. The culture medium is clear in components, high in cultured cell density, free of animal serum, beneficial to purification of downstream products, capable of improving product quality, convenient to prepare and use, and suitable for full-suspension serum-free culture of DF-1 cells and large-scale production of chicken bursal disease vaccines.

Description

technical field [0001] The invention belongs to the technical field of cell serum-free medium, and in particular relates to a serum-free medium for cultivating DF-1 cells and a preparation method thereof. Background technique [0002] DF-1 cell is a duck fibroblast cell line, which is widely used as an immortalized cell line for culturing and propagating chicken bursal virus. Chicken bursal virus infection is a viral infectious disease mainly caused by broiler chickens. Since 2015, the disease has been widely prevalent in China, and the lethality rate of 3-5 week-old broilers is as high as 40% to 100%, which has caused huge economic losses to the poultry industry in my country. [0003] At present, the most effective preventive measures are vaccines. The key to preparing high-efficiency vaccines is to obtain high-titer viruses. Traditional vaccine production is to culture DF-1 cells on the wall with serum-containing medium and then inoculate them. The cell concentration is...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N7/00C12R1/93
CPCC12N5/0656C12N7/00C12N2510/04C12N2500/90C12N2500/32C12N2500/38C12N2500/16C12N2500/14C12N2500/12C12N2500/30C12N2500/36C12N2500/20C12N2500/24C12N2500/60C12N2500/76C12N2500/34C12N2500/40C12N2720/10051
Inventor 王洪林刘志亮孙友君范娜娜宋桂才张恒卢博王稼林马广斌鹿洪伟王月华邹海涛
Owner 山东信得动物疫苗有限公司
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