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Serum-free culture medium for adoptive immune cells

A technology of serum-free medium and immune cells, applied in the field of cell engineering, to achieve stable product performance, promote the formation of lentivirus-infected primary T lymphocytes and carT cells, and have strong reproducibility

Inactive Publication Date: 2016-12-07
李昂 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the incorporation of human serum brings the risk of HIV, HBV and other viral infections

Method used

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  • Serum-free culture medium for adoptive immune cells
  • Serum-free culture medium for adoptive immune cells
  • Serum-free culture medium for adoptive immune cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] T cell culture

[0022] The formulation of the serum-free medium for adoptive immune cells provided by this embodiment is as follows:

[0023] RPMI 1640 medium, and additional nutritional components added to the RPMI 1640 medium, the types of nutritional components and the added quality per unit volume are as follows: L-arginine 156mg / L, L-asparagine 53mg / L, L-Aspartic Acid 17mg / L, L-Glutamic Acid 19mg / L, L-Glutamine 285mg / L, Glycine 17mg / L, L-Histidine 23.5mg / L, L-Isoleucine 53mg / L, L-polylysine sodium 33mg / L, L-methionine 23.5mg / L, L-phenylalanine 21mg / L, L-proline 19mg / L, glucose 2500mg / L, L-serine 28mg / L, L-threonine 20.5mg / L, L-tryptophan 11mg / L, L-valine 23.5mg / L, L-leucine 53mg / L, β-mercaptoethanol 3.2mg / L , L-tyrosine disodium dihydrate 304mg / L, ethanolamine 2.5mg / L, L-cystine dihydrochloride 53mg / L, dexamethasone 0.002mg / L, CuSO 4 ·5H 2 O0.0009mg / L, MnCl 2 4H 2 O 0.00045mg / L, Ni(NO3) 2 ·6H 2 O 0.00007mg / L, ZnSO 4 ·7H 2 O 0.19mg / L, CoCl 2 ·6H 2 O 0...

Embodiment 2

[0032] T cell culture

[0033] The formulation of the serum-free medium for adoptive immune cells provided by this embodiment is as follows:

[0034] Get the RPMI 1640 culture medium of unit volume, add the nutrient component in the RPMI 1640 culture medium unit volume addition amount as follows: L-arginine 165mg / L, L-asparagine 56mg / L, L-asparagine Acid 18mg / L, L-glutamic acid 20mg / L, L-glutamine 300mg / L, glycine 18mg / L, L-histidine 25mg / L, L-isoleucine 56mg / L, L-poly Sodium polylysine 35mg / L, L-methionine 25mg / L, L-phenylalanine 23mg / L, L-proline 20mg / L, glucose 2500-3000mg / L, L-serine 30mg / L, L - Threonine 22mg / L, L-Tryptophan 12mg / L, L-Valine 25mg / L, L-Leucine 55mg / L, β-Mercaptoethanol 3.5mg / L, L-Tyramine Dihydrate Disodium acid 320mg / L, ethanolamine 3mg / L, L-cystine dihydrochloride 55mg / L, dexamethasone 0.003mg / L, CuSO 4 5H2O 0.001mg / L, MnCl 2 4H 2 O 0.0005mg / L, Ni(NO3) 2 ·6H 2 O 0.00008mg / L, ZnSO 4 ·7H 2 O 0.2mg / L, CoCl 2 ·6H 2 O 0.005mg / L, SnC1 2 2H 2 O0.0...

Embodiment 3

[0043] T cell culture

[0044] RPMI 1640 medium, and additional nutritional components added to the RPMI 1640 medium, the types of nutritional components and the added quality per unit volume are as follows: L-arginine 174mg / L, L-asparagine 59mg / L, L-aspartic acid 19mg / L, L-glutamic acid 21mg / L, L-glutamine 315mg / L, glycine 19mg / L, L-histidine 26.5mg / L, L-iso Leucine 59mg / L, L-polylysine sodium 37mg / L, L-methionine 26.5mg / L, L-phenylalanine 24.5mg / L, L-proline 21mg / L, glucose 3000mg / L L, L-serine 32mg / L, L-threonine 23mg / L, L-tryptophan 13mg / L, L-valine 26.5mg / L, L-leucine 58mg / L, β-mercaptoethanol 3.7mg / L, L-tyrosine disodium dihydrate 336mg / L, ethanolamine 3.2mg / L, L-cystine dihydrochloride 58mg / L, dexamethasone 0.004mg / L, CuSO 4 ·5H 2 O 0.0011mg / L, MnCl 2 4H 2 O 0.00055mg / L, Ni(NO3) 2 ·6H 2 O0.00009mg / L, ZnSO 4 ·7H 2 O 0.21mg / L, CoCl 2 ·6H 2 O 0.006mg / L, SnC1 2 2H 2 O 0.00007 mg / L, Na 2 SeO 3 0.01mg / L,FeSO 4 ·7H 2 O 1.3mg / L, vitamin C 0.55mg / L, p-hydroxyb...

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Abstract

The present invention provides a serum-free culture medium for adoptive immune cells. The serum-free culture medium is characterized by comprising: a RPMI 1640 culture medium and nutrition components additionally added to the RPMI 1640 culture medium, wherein the nutrition components comprise L-arginine, L-asparagine, L-aspartic acid, L-glutamic acid, L-glutamine, glycine, L-histidine, L-isoleucine, L-polylysine sodium, L-methionine, L-phenylalanine, L-proline, glucose, L-serine, L-threonine, L-tryptophan, L-valine, L-leucine, beta-mercaptoethanol, L-tyrosine disodium dihydrate, ethanolamine, L-cystine dihydrochloride, dexamethasone and the like. The serum-free culture medium of the present invention has no toxicity and is beneficial for the subsequent clinical application.

Description

technical field [0001] The invention relates to a serum-free culture medium for adoptive immune cells, belonging to the field of cell engineering. Background technique [0002] Adoptive immune cell therapy is considered to be the fourth tumor treatment after surgery, chemotherapy and radiotherapy. More and more studies have shown that adoptive immune cell therapy will be the main treatment method to overcome tumors in the future. In 2013, the emergence of genetically modified T cell (carT) therapy pushed tumor immune cell therapy to the peak. Science magazine listed carT therapy as the top ten scientific discoveries of the year. At present, many pharmaceutical giants at home and abroad have invested heavily in the research and development of carT therapy for various tumors. It is predicted that by 2025, the market size of carT and related immune cell therapies will reach 30-40 billion US dollars. [0003] The key to carT and related immune cell therapy is to expand the c...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0784
Inventor 李昂刘荣军
Owner 李昂
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