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Serum-free culture medium suitable for immune cell large-scale culture

A serum-free medium, large-scale culture technology, applied in the field of cell biology and medical immunology, can solve the problems of difficult to meet the reinfusion standard, long culture time, etc., to avoid safety problems, reduce production costs, and increase proliferation effect. Good results

Active Publication Date: 2015-11-11
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the most widely used serum-free medium is LONZA’s XVIVO serum-free medium and GIBCO’s AIM serum-free medium, but the above two mediums need to be supplemented with autologous plasma in the early stage of cell culture, otherwise, when culturing immune cells, It is difficult to meet the clinical reinfusion standard, and the culture time is long

Method used

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  • Serum-free culture medium suitable for immune cell large-scale culture
  • Serum-free culture medium suitable for immune cell large-scale culture
  • Serum-free culture medium suitable for immune cell large-scale culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A serum-free medium suitable for large-scale cultivation of immune cells, based on RPMI1640 medium, also contains additional components, the concentration of which is: hydroxyethylpiperazine sulfuric acid 3 / L, phenol red sodium 0.0001g / L, L-glutamate 0.1g / L, sodium bicarbonate 0.5g / L. The added ingredients also include 0.001g / L of insulin, 0.001g / L of transferrin, 0.0001g / L of selenium, 2g / L of human albumin, and 240IU / mL of interleukin.

[0043] The RPMI1640 medium includes inorganic salts, amino acids, vitamins, and sugars.

[0044] The concentration of the inorganic salt is: calcium dinitrate tetrahydrate 0.01g / L, magnesium sulfate anhydrous 0.04g / L, potassium chloride 0.04g / L, sodium bicarbonate 0.2g / L, sodium chloride 2g / L, Disodium hydrogen phosphate 0.08g / L; The concentration of the amino acid is: L-arginine

[0045] 0.02g / L, L-valine 0.002g / L, L-aspartic acid 0.002g / L, L-asparagine 0.005g / L, L-cysteine ​​dihydrochloride 0.006g / L, L-glutamic acid 0.002g / L, L-g...

Embodiment 2

[0051] A serum-free medium suitable for large-scale cultivation of immune cells, based on RPMI1640 medium, also contains additional components, the concentration of which is: hydroxyethylpiperazine sulfuric acid 9 / L, phenol red sodium 0.1g / L, L-glutamate 1g / L, sodium bicarbonate 8g / L. The added ingredients also include insulin 0.1g / L, transferrin 0.01g / L, selenium 0.1g / L, human albumin 60g / L, interleukin-2600IU / mL.

[0052] The RPMI1640 medium includes inorganic salts, amino acids, vitamins, and sugars.

[0053] The concentration of the inorganic salt is: calcium dinitrate tetrahydrate 1g / L, magnesium sulfate anhydrous 0.4g / L, potassium chloride 2g / L, sodium bicarbonate 8g / L, sodium chloride 10g / L, dihydrogen phosphate Sodium 4g / L; the concentration of the amino acid is: L-arginine 2g / L, L-valine 2g / L, L-aspartic acid 2g / L, L-asparagine 0.5g / L, L-cysteine·dihydrochloride 0.6g / L, L-glutamic acid 0.2g / L, L-glutamine 0.3g / L, L-histidine 0.15g / L, glycine 0.1g / L, L-isoleucine 0....

Embodiment 3

[0059] A serum-free medium suitable for large-scale cultivation of immune cells, based on RPMI1640 medium, also contains additional components, the concentration of which is: hydroxyethylpiperazine sulfuric acid 5g / L, phenol red sodium 0.002g / L, L-glutamate 0.2g / L, sodium bicarbonate 1g / L, insulin 0.005g / L, transferrin 0.002g / L, selenium 0.001g / L, human albumin 5g / L , Interleukin-250IU / mL.

[0060] The RPMI1640 medium includes inorganic salts, amino acids, vitamins, and sugars.

[0061] The concentration of the inorganic salt is: calcium dinitrate tetrahydrate 0.1g / L, magnesium sulfate anhydrous 0.04884g / L, potassium chloride 0.4g / L, sodium bicarbonate 2g / L, sodium chloride 6g / L, phosphoric acid Disodium hydrogen 0.8g / L; The concentration of the amino acid is: L-arginine 0.2g / L, L-valine 0.02g / L, L-aspartic acid 0.02g / L, L-aspartic acid Amide 0.05g / L, L-cysteine·dihydrochloride 0.0652g / L, L-glutamic acid 0.02g / L, L-glutamine 0.3g / L, L-histidine 0.015g / L, Glycine 0.01g / L, L-...

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Abstract

The invention discloses a serum-free culture medium suitable for immune cell large-scale culture, and belongs to the field of cell biology and medical immunology. The serum-free culture medium takes an RPMI1640 culture medium as a basis, additive components are further contained and include HEPES, phenol red sodium salt, L-glutamate and sodium bicarbonate. Insulin, transferrin, selenium, human albumin and interleukin-2 can be further added. Components contained in the culture medium are chemical defined components, no animal serum components are contained, and the serum-free culture medium is suitable for large-scale culture of immunity cells and is high in clinical safety and good in proliferation effect.

Description

technical field [0001] The invention belongs to the fields of cell biology and medical immunology, and in particular relates to a serum-free medium suitable for culturing all immune cells. Background technique [0002] With the rapid development of immune cell biology and immune molecular biology, somatic cell immunotherapy has become one of the important means of adjuvant therapy for tumor patients after radiotherapy and chemotherapy. All have good effect. [0003] Somatic cell immunotherapy is a biological therapy that injects immune cells with anti-tumor activity into tumor patients to directly kill the tumor or stimulate the body's anti-tumor immune response to achieve therapeutic effects. In vitro manipulations include cell passage, expansion, modification, screening and treatment with drugs or other treatments that can change the biological behavior of cells in vitro. Somatic cells manipulated in vitro can be used for tumor treatment and also for tumor prevention. ...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0784
Inventor 葛啸虎陈海佳王一飞罗二梅王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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