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High efficiency fetal calf serum substitute

A technology of fetal bovine serum and substitutes, applied in the field of serum substitutes, can solve the problems that affect the widespread use of serum substitutes, increase the risk of spreading diseases, spread infectious diseases, etc., and achieve the benefits of large-scale use and avoid the spread of infectious diseases risk, the effect of increasing supply

Inactive Publication Date: 2015-08-12
上海泰坦科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cellular immunotherapy is inseparable from the cultivation of a large number of lymphocytes in vitro, and the cultivation of lymphocytes requires a large amount of culture medium. The advantages of using animal serum are sufficient supply and low price, but the disadvantage of using animal serum is the potential transmission of infection risk of disease, and the development of allergies due to foreign proteins
[0003] CN102352343A (published on February 15, 2012) discloses a lymphocyte culture medium, including a basal medium and a serum substitute, and the serum substitute includes recombinant human serum albumin, recombinant human transferrin, recombinant human insulin , lipids, trace element compounds and vitamins to solve the problem that most of the existing lymphocyte culture media use animal or human serum and its components as additives, which increases the risk of spreading diseases
[0004] Although the above-mentioned prior art discloses some fetal bovine serum substitutes, which can meet certain needs, there are still certain defects in these: for example, the survival rate of lymphocytes cultured in vitro is only about 92%, and the value-added multiple is only about 22, which is obviously low. in the control group, which would affect the widespread use of the above-mentioned serum replacement

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1, the configuration of lymphocyte culture medium

[0014] Add 2mg of recombinant human serum albumin, 50mg of recombinant human transferrin, 19mg of galactose, 2mg of cholesterol, 0.2mg of linoleic acid, 0.2mg of linolenic acid, 1mg of vitamin E, 802mg of Tween, and iron nitrate into each 1L of RPMI 1640 medium 0.1mg, sodium selenite 0.01mg, copper sulfate 0.001mg, zinc sulfate 0.5mg, ethanolamine 1.5mg, ascorbyl phosphate 2.5mg, gentamicin 10IU, stir to fully dissolve, add 0.1mol / L hydrochloric acid and 0.1mol / L sodium hydroxide to adjust the pH value between 6.8, filter and sterilize with a 0.1um filter membrane, pack in aliquots, and add 300 mg of glutamine just before use.

Embodiment 2

[0015] Embodiment 2, the configuration of lymphocyte culture medium

[0016] Add 2mg of recombinant human serum albumin, 50mg of recombinant human transferrin, 15mg of galactose, 2mg of cholesterol, 0.2mg of linoleic acid, 0.2mg of linolenic acid, 1mg of vitamin E, 802mg of Tween, and iron nitrate into each 1L of RPMI 1640 medium 0.1mg, sodium selenite 0.01mg, copper sulfate 0.001mg, zinc sulfate 0.5mg, ethanolamine 1.5mg, ascorbyl phosphate 2.5mg, gentamicin 10IU, stir to fully dissolve, add 0.1mol / L hydrochloric acid and 0.1mol / L sodium hydroxide to adjust the pH value between 7.0, 0.1um filter membrane filter sterilization, subpackage, add glutamine 300mg before use.

Embodiment 3

[0017] Embodiment 3, the configuration of lymphocyte culture medium

[0018] Add 2 mg of recombinant human serum albumin, 50 mg of recombinant human transferrin, 25 mg of galactose, 2 mg of cholesterol, 0.2 mg of linoleic acid, 0.2 mg of linolenic acid, 1 mg of vitamin E, 802 mg of Tween, and iron nitrate into each 1 L of RPMI 1640 medium 0.1mg, sodium selenite 0.01mg, copper sulfate 0.001mg, zinc sulfate 0.5mg, ethanolamine 1.5mg, ascorbyl phosphate 2.5mg, gentamicin 10IU, stir to fully dissolve, add 0.1mol / L hydrochloric acid and 0.1mol / L sodium hydroxide to adjust the pH value between 7.2, 0.1um filter membrane filter sterilization, sub-package, add glutamine 300mg before use.

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PUM

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Abstract

The invention relates to a high efficiency fetal calf serum substitute. The high efficiency fetal calf serum substitute is composed of recombinant human albumin, recombinant human transferrin, galactose, lipid, a trace element compound and vitamins, a weight ratio of the recombinant human albumin to the recombinant human transferrin to the galactose is 2:50:(15-25), and the lipid is a cholesterol, linoleic acid and linolenic acid mixture. The fetal calf serum substitute provided by the invention avoids infectious disease propagation risk and foreign protein induced allergy, increases the in vitro culture survival rate and the increment multiple of human lymphocytes to 96% and 28 respectively, improves the supply of the lymphocytes, reduces the culture cost, and facilitates large scale use.

Description

technical field [0001] The invention relates to a serum substitute, in particular to a high-efficiency fetal bovine serum substitute. Background technique [0002] The medium provides basic environmental and nutrient factors for cell growth, including osmotic pressure, pH value, inorganic salts, amino acids, functional proteins, vitamins, etc. In order to provide these nutritional factors, traditional culture media mostly use animal or human serum and its components as additives. The more common ones include newborn bovine serum, fetal bovine serum, human AB serum, bovine serum albumin, and human serum albumin. , human transferrin, etc. Cellular immunotherapy is inseparable from the cultivation of a large number of lymphocytes in vitro, and the cultivation of lymphocytes requires a large amount of culture medium. The advantages of using animal serum are sufficient supply and low price, but the disadvantage of using animal serum is the potential transmission of infection ri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/0783
Inventor 谢应波张庆张华徐肖冰罗桂云
Owner 上海泰坦科技股份有限公司
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