Serum-free medium of peripheral blood cells

A technology of serum-free culture medium and culture medium, applied in the field of cell culture, which can solve the problems of virus contamination, inaccurate test data, interference of bovine serum components in experimental research, etc., and achieve good proliferation effect and good cell survival rate

Inactive Publication Date: 2017-08-15
SHANGHAI XP BIOMED
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] The currently commonly used culture medium for peripheral blood cells adopts the traditional bovine serum-containing medium. However, the high cost of bovine serum, the risk of virus contamination, and the interference of bovine serum components with experimental research may lead to inaccurate test data. Therefore, serum-free medium And serum-free culture is a major trend in cell culture

Method used

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  • Serum-free medium of peripheral blood cells
  • Serum-free medium of peripheral blood cells
  • Serum-free medium of peripheral blood cells

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Effect test

Embodiment 1

[0022] The preparation process of the serum-free medium of the peripheral blood cells of embodiment 1 is as follows:

[0023] Step 1): Weigh 168 g of the synthesized RPMI 1640 dry powder on an electronic balance under sterile conditions, place it in a sterile container, add 9 L of purified water to dilute, and stir evenly to obtain a basic medium.

[0024] Step 2): add components: 32g sodium bicarbonate, 85g L-glutamine, 420mg interleukin-2, 12.6g heparin, 280mg transferrin, 270mg human recombinant insulin, 155mg M-type phytohemagglutinin, 2.5g gentamicin (equivalent to 2.5×10 5 IU / L) and 3g streptomycin (equivalent to 3×10 5 IU / L), and 70 mg of phenol red sodium salt were sequentially added to the basal medium obtained in step 1), and thoroughly mixed.

[0025] Step 3): Sterilize by filtration, then adjust the pH value to 7.4±0.2, and finally add purified water to adjust the volume to 10L;

[0026] Step 4): After the culture medium is prepared, it is sterilized by filtrati...

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Abstract

The invention relates to a serum-free medium of peripheral blood cells. The serum-free medium comprises a base medium and an additive component; the base medium is RPMI 1640 medium; the additive component is composed of, by volume, 35 to 50mg/L of interleukin-2, 1 to 2g/L of heparin, 25 to 30mg/L of transferring, 20 to 30mg/L of insulin human recombinant, 10 to 20mg/L of M-type phytohaemagglutinin, 7 to 10mg/L of L-glutamine, and 2 to 4g/L of sodium bicarbonate. Compared with the prior art, the serum-free medium of peripheral blood cells possesses following advantages: the serum-free medium of peripheral blood cells is capable of satisfying requirements of leukocyte rapid and large amount propagation in peripheral blood cells, is especially suitable to be used for culturing of peripheral blood cells, and possesses excellent propagation effect and cell survival rate.

Description

technical field [0001] The invention relates to a serum-free medium for peripheral blood cells, belonging to the technical field of cell culture. Background technique [0002] Cell culture has been widely used in various fields such as biology, medicine, and new drug research and development, and has become one of the most important basic sciences. [0003] Human peripheral blood cell culture is widely used in medical testing, especially in genetics, and is one of the important basic testing techniques. [0004] The currently commonly used culture medium for peripheral blood cells adopts the traditional bovine serum-containing medium. However, the high cost of bovine serum, the risk of virus contamination, and the interference of bovine serum components with experimental research may lead to inaccurate test data. Therefore, serum-free medium And serum-free culture is a major trend in cell culture. [0005] However, after adopting the serum-free medium, which serum replacem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2500/25C12N2500/32C12N2500/90C12N2501/2302C12N2501/59C12N2501/91
Inventor 颜学恒苏恩筠张智培薛宜青熊延狮
Owner SHANGHAI XP BIOMED
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