A kind of NK cell serum-free medium and its application

A serum-free medium, NK cell technology, applied in the field of NK cell serum-free medium, can solve the problems of insufficient amplification efficiency and cell quality to be improved, and achieves high viability rate, few types of added components, and improved lethality. Effect

Active Publication Date: 2022-01-07
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, isolated human plasma needs to be added when culturing cells, and the amplification efficiency is slightly insufficient, and the cell quality needs to be improved

Method used

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  • A kind of NK cell serum-free medium and its application
  • A kind of NK cell serum-free medium and its application
  • A kind of NK cell serum-free medium and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A serum-free culture medium for NK cells, the culture medium includes a basal medium and additional components, the basal medium is an IMDM medium, and the volume ratio of the basal medium to the additional components is 1000mL: 20mL.

[0034]Based on the volume of the added components, the content of each component of the added components is as follows: rhIL-2 (inshore protein, product number GMP-CD66) 20 ng / mL, rhIL-15 (inshore protein, product number GMP-CD66) 20 ng / mL, rhIL-15 (inshore protein, product number GMP- CD66) 10ng / mL, rhIL-21 (nearshore protein, product number GMP-CD66) 0.1ng / mL, FK506 (MedChemExpress, product number HY-13756) 10ng / mL, mevalonate (Sigma, product number 90469) 0.5µg / mL mL, GGPP (Sigma, Cat. No. G6025) 10 µg / mL, recombinant human insulin (Sigma, Cat. No. 91077c) 0.01 µg / mL, recombinant apotransferrin (Sigma, Cat. No. T1147) 10 µg / mL, recombinant human serum albumin (Heyuan Biology, product number HYC002M01) 5 mg / mL, sodium pyruvate (Sigma, ...

Embodiment 2

[0036] A serum-free culture medium for NK cells, the culture medium includes a basal medium and additional components, the basal medium is an IMDM medium, and the volume ratio of the basal medium to the additional components is 1000mL: 20mL.

[0037] Based on the volume of the added components, the content of each component of the added components is as follows: rhIL-2 (nearshore protein, product number GMP-CD66) 5ng / mL, rhIL-15 (nearshore protein, product number GMP-CD66 ) 20ng / mL, rhIL-21 (nearshore protein, product number GMP-CD66) 5 ng / mL, FK506 (MedChemExpress, product number HY-13756) 50 ng / mL, mevalonate (Sigma, product number 90469) 1.5 µg / mL mL, GGPP (Sigma, catalog number G6025) 10 µg / mL, recombinant human insulin (Sigma, catalog number 91077c) 10 µg / mL, recombinant apotransferrin (Sigma, catalog number T1147) 5 µg / mL, recombinant human serum albumin ( Heyuan Bio, Product No. HYC002M01) 2 mg / mL, Sodium Pyruvate (Sigma, Product No. P5280) 50 µg / mL, Linoleic Acid (Sigm...

Embodiment 3

[0039] A serum-free culture medium for NK cells, the culture medium includes a basal medium and additional components, the basal medium is an IMDM medium, and the volume ratio of the basal medium to the additional components is 1000mL: 20mL.

[0040] Based on the volume of the added components, the content of each component of the added components is as follows: rhIL-2 (nearshore protein, product number GMP-CD66) 0.1ng / mL, rhIL-15 (nearshore protein, product number GMP-CD66) CD66) 100 ng / mL, rhIL-21 (Lisshore Protein, Cat. No. GMP-CD66) 20 ng / mL, FK506 (MedChemExpress, Cat. No. HY-13756) 100 ng / mL, mevalonate (Sigma, Cat. No. 90469)5 µg / mL, GGPP (Sigma, Cat. No. G6025) 2 µg / mL, recombinant human insulin (Sigma, Cat. No. 91077c) 15 µg / mL, recombinant apotransferrin (Sigma, Cat. No. T1147) 0.1 µg / mL, recombinant human serum Albumin (Heyuan Bio, Product No. HYC002M01) 0.1 mg / mL, sodium pyruvate (Sigma, Product No. P5280) 100 µg / mL, linoleic acid (Sigma, Product No. L1012) 10 µg / m...

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Abstract

The invention discloses a serum-free culture medium for NK cells and an application thereof. The serum-free culture medium for NK cells is prepared by mixing a basal culture medium and additional components, and the additional components are composed of rhIL-2, rhIL-15 , rhIL‑21, tacrolimus, mevalonate, isoprene pyrophosphate, recombinant human insulin, recombinant human apotransferrin, recombinant human serum albumin, sodium pyruvate, linoleic acid, cholesterol , composed of folinic acid, and the basal medium is IMDM. The culture medium of the present invention not only has a clear chemical composition, is serum-free, has no source and animal source components, has a simple formula, and has excellent performance. At the same time, FK506 is added to the serum-free medium, which can significantly improve NK cell activity while ensuring high cell viability. The expansion speed of NK cells can be expanded by more than 700 times after 17 days of culture, and the cell viability can reach more than 95% when harvested.

Description

technical field [0001] The invention relates to the technical field of culture medium, in particular to a serum-free culture medium for NK cells and its application. Background technique [0002] Natural killer cells, namely NK cells (Natural Killer Cells), are a type of lymphocytes with the same name as T cells and B cells but different from them. At the same time, they have a certain regulatory effect on T lymphocytes and B lymphocytes in the human body. . NK cells are a type of immune cells that have nothing to do with specific immune responses. The cell lysis mediated by them is not affected by the major histocompatibility complex (MHC), so they are also called the "heart" of the body's natural immune system. [0003] As the body's first line of defense, NK cells can not only clear intracellular parasites, viruses, and aging mutant cells, but also have a strong clearing effect on cancer cells, and can be rapidly activated without prior sensitization. It has powerful an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2302C12N2501/2315C12N2501/2321C12N2500/25C12N2500/90C12N2501/998C12N2500/36C12N2500/30C12N2500/38C12N2501/04
Inventor 林家会马士棋陈旭陈刚
Owner 苏州依科赛生物科技股份有限公司
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