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A kind of immune cell serum-free culture medium and its application

A technology of immune cells and culture medium, which is applied in the fields of medicine and biology, can solve the problems of long doubling time, slow cell proliferation, and inability to mass-produce, and achieve good stability

Active Publication Date: 2018-08-28
友康厚德生物制品(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method has slow cell proliferation, long doubling time, and CD3 cells + CD56 + Positive ratio is unstable
And because the cell culture medium containing animal or human serum is used for cultivation, and the animal or human serum contains pathogens or uncertain substances, which will bring dangerous or uncertain harm to the cultured cells
Moreover, the traditional medium components are unstable and cannot be used for quality control in the later stage, and cannot be mass-produced in industry

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Weigh the reagents according to the following ratio, and the final concentration unit is mg / L: L-arginine 300, CuSO 4 ·5H 2 O0.005, L-asparagine 75, L-aspartic acid 30, L-glutamic acid 30, Ni(NO 3 ) 2 ·6H 2 O 0.0002, L-Glutamine 500, ZnSO 4 ·7H 2 O 0.6, Glycine 15, CoCl 2 ·6H 2 O 0.008, L-histidine 30, NaSiO 3 9H 2 O 0.01, L-isoleucine 27, Na 3 VO 4· 12H 2 O 0.005, L-lysine hydrochloride 60, SnC1 2 2H 2 O 0.0001, L-methionine 30, Na 2 SeO 3 0.01, L-phenylalanine 30, FeSO 4 ·7H 2 O 1.6, L-proline 30, glucose 4000, L-serine 45, vitamin C 0.704, L-threonine 30, p-hydroxybenzoic acid 1.5, L-tryptophan 15, sodium pyruvate 550, L-valeric acid Amino acid 30, linoleic acid 0.05, L-leucine 75, β-mercaptoethanol 4.0, L-tyrosine disodium dihydrate 432, ethanolamine 5, L-cystine dihydrochloride 75, dexamethasone 0.005, Human transferrin 50, insulin 20, purified human transferrin, cholesterol 60, catalase 50, sodium selenite 35 and 1% dimercaptoethanol solution 1....

Embodiment 2

[0019] Weigh the reagents according to the following ratio, and the final concentration unit is mg / L:

[0020] L-Arginine 100, CuSO 4 ·5H 2 O0.0005, L-asparagine 25, L-aspartic acid 10, L-glutamic acid 10, Ni(NO 3 ) 2 ·6H 2 O 0.00002, L-Glutamine 200, ZnSO 4 ·7H 2 O0.06, Glycine 5, CoCl 2 ·6H 2 O0.001, L-histidine 10, NaSiO 3 9H 2 O0.001, L-isoleucine 25, Na 3 VO 4 12H 2 O 0.0005, L-lysine hydrochloride 20, SnC1 2 2H 2 O 0.00001, L-methionine 10, Na2 SeO 3 0.002, L-phenylalanine 10, FeSO 4 ·7H 2 O0.2, L-proline 10, glucose 1000, L-serine 15, vitamin C 0.176, L-threonine 10, p-hydroxybenzoic acid 0.5, L-tryptophan 5, sodium pyruvate 55, L- Valine 10, linoleic acid 0.01, L-leucine 25, β-mercaptoethanol 0.8, L-tyrosine disodium dihydrate 144, ethanolamine 1, L-cystine dihydrochloride 25, dexamethasone 0.001 , human transferrin 5, insulin 8, purified human transferrin 1, cholesterol 20, catalase 20, sodium selenite 17.3 and 1% dimercaptoethanol solution 0.35ml / L...

Embodiment 3

[0022] Weigh the reagents according to the following ratio, and the final concentration unit is mg / L:

[0023] L-Arginine 150, CuSO 4 ·5H 2 O0.001, L-asparagine 50, L-aspartic acid 20, L-glutamic acid 20, Ni(NO 3 ) 2 ·6H 2 O 0.0001, L-Glutamine 300, ZnSO 4 ·7H 2 O0.03, Glycine 10, CoCl 2 ·6H 2 O 0.004, L-histidine 20, NaSiO 3 9H 2 O0.008, L-isoleucine 26, Na 3 VO 4 12H 2 O 0.0001, L-lysine hydrochloride 30, SnC1 2 2H 2 O 0.00008, L-methionine 20, Na 2 SeO 3 0.008, L-phenylalanine 20, FeSO 4 ·7H 2 O1.0, L-proline 20, glucose 2000, L-serine 30, vitamin C 0.374, L-threonine 20, p-hydroxybenzoic acid 1.0, L-tryptophan 10, sodium pyruvate 350, L- Valine 20, linoleic acid 0.04, L-leucine 50, β-mercaptoethanol 2.3, L-tyrosine disodium dihydrate 319, ethanolamine 4, L-cystine dihydrochloride 50, dexamethasone 0.003 , human transferrin 25, insulin 10, purified human transferrin 5, cholesterol 40, catalase 30, sodium selenite 17.3, and 1% dimercaptoethanol solution 0...

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Abstract

The invention relates to the field of biology and medicine, in particular to an immune cell serum-free medium and preparation and application thereof. The serum-free medium is prepared from dexamethasone, human transferrin, insulin, cholesterol, catalase, sodium selenite and mercaptoethanol. The serum-free medium is safe to the human body, and can increase the CIK cell proliferation speed and stabilize cells CD3+CD56+, and the serum-free medium has the advantages that the components are simple, the character is stable, the CIK proliferation induction efficiency is high, the positive proportion of CD3+CD56+ is high, and the quality of CIK cells obtained through culture is stable.

Description

technical field [0001] The invention relates to the fields of biology and medicine, in particular to a serum-free culture medium of immune cells and its application. Background technique [0002] Tumor adoptive immunotherapy (ACI) refers to the infusion of immune cells with anti-tumor activity into tumor patients to directly kill or stimulate the body's immune response to kill tumor cells, so as to achieve the purpose of treating tumors. It includes non-specifically activated and specifically activated effector cells. The former uses non-specific stimulating factors (IL-2, interferon) to stimulate precursor effector cells to activate them into effector cells with anti-tumor activity, such as LAK cells, Tumor infiltrating lymphocytes (TIL), cytokine-induced killer cells (Cytokine induced killer cells, CIK), etc.; specifically activated effector cells refer to anti-tumor effector cells induced by using tumor antigens as stimuli, such as dendrites Cytotoxic T lymphocytes (CD8 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078C12N5/0783
Inventor 曲宝赤
Owner 友康厚德生物制品(北京)有限公司