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Serum-free culture medium for amplification in vitro of human immune killer cell

A serum-free medium and cell-killing technology, applied in the field of medium research and development of modern biotechnology, can solve the problem of increasing heterologous infection

Active Publication Date: 2017-06-13
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, conventional cell culture is carried out in a system with serum, and the addition of heterologous serum may increase the risk of heterologous infection, which is absolutely not allowed in clinical practice

Method used

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  • Serum-free culture medium for amplification in vitro of human immune killer cell
  • Serum-free culture medium for amplification in vitro of human immune killer cell
  • Serum-free culture medium for amplification in vitro of human immune killer cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0164] (1) A basal medium for the in vitro expansion of human immune killer cells, named BM1, its various components and final use concentrations are (unit: mg / L):

[0165] Calcium Chloride CaCl 2 0.1;

[0166] Calcium nitrate Ca(NO 3 ) 2 69.5;

[0167] Potassium chloride KCl 400

[0168] Potassium nitrate KNO 3 0.001;

[0169] Disodium hydrogen phosphate Na 2 HPO 4 423.88;

[0170] Sodium chloride NaCl 6000;

[0171] Sodium dihydrogen phosphate monohydrate NaH 2 PO4·H 2 O 676.13;

[0172] Magnesium Sulfate MgSO 4 40.276;

[0173] Magnesium Chloride MgCl 2 0.1;

[0174] Sodium bicarbonate NaHCO 3 2000;

[0175] Sodium Selenite Na 2 SeO 3 0.001;

[0176] Iron nitrate nonahydrate Fe (NO 3 ) 3 9H 2 O 0.001;

[0177] Heptahydrate ferrous sulfate FeSO 3 ·7H 2 O 0.001;

[0178] Copper Sulfate Pentahydrate CuSO 4 ·5H 2 O 0.00001;

[0179] Zinc sulfate heptahydrate ZnSO 4 ·7H 2 O 0.001;

[0180] L-Alanine L-Alanine 0.1;

[0181] L-arginine·hy...

Embodiment 2

[0237] (1) A basal medium for the in vitro expansion of human immune killer cells, named BM2, its various components and final use concentrations are (unit: mg / L):

[0238] Calcium Chloride CaCl 2 165;

[0239] Calcium nitrate Ca(NO 3 ) 2 0.1;

[0240] Potassium chloride KCl 320

[0241] Potassium nitrate KNO 3 0.038

[0242] Disodium hydrogen phosphate Na 2 HPO 4 0.1;

[0243] Sodium chloride NaCl 5747.75;

[0244] Sodium dihydrogen phosphate monohydrate NaH 2 PO4·H 2 O 93.75;

[0245] Magnesium Sulfate MgSO 4 73.255;

[0246] Magnesium Chloride MgCl 2 14.32;

[0247] Sodium bicarbonate NaHCO 3 2731;

[0248] Sodium Selenite Na 2 SeO 3 0.0114;

[0249] Iron nitrate nonahydrate Fe (NO 3 ) 3 9H 2 O 0.025;

[0250] Heptahydrate ferrous sulfate FeSO 3 ·7H 2 O 0.2085;

[0251] Copper Sulfate Pentahydrate CuSO 4 ·5H 2 O 0.00065;

[0252] Zinc sulfate heptahydrate ZnSO 4 ·7H 2 O 0.216;

[0253] L-Alanine L-Alanine 14.725;

[0254] L-Argini...

Embodiment 3

[0318] (1) A basal medium for the in vitro expansion of human immune killer cells, named BM3, its various components and final use concentrations are (unit: mg / L):

[0319] Calcium Chloride CaCl 2 116.6;

[0320] Calcium nitrate Ca(NO 3 ) 2 0.1;

[0321] Potassium chloride KCl 311.8;

[0322] Potassium nitrate KNO 3 0.076;

[0323] Disodium hydrogen phosphate Na 2 HPO 4 71.02;

[0324] Sodium chloride NaCl 6995.5;

[0325] Sodium dihydrogen phosphate monohydrate NaH 2 PO4·H 2 O 62.5;

[0326] Magnesium Sulfate MgSO 4 48.84;

[0327] Magnesium Chloride MgCl 2 28.64;

[0328] Sodium bicarbonate NaHCO 3 2438;

[0329] Sodium Selenite Na 2 SeO 3 0.0001;

[0330] Iron nitrate nonahydrate Fe (NO 3 ) 3 9H 2 O 0.05;

[0331] Heptahydrate ferrous sulfate FeSO 3 ·7H 2 O 0.417;

[0332] Copper Sulfate Pentahydrate CuSO 4 ·5H 2 O 0.0013;

[0333] Zinc sulfate heptahydrate ZnSO 4 ·7H 2 O 0.432;

[0334] L-Alanine L-Alanine 4.45;

[0335] L-Arginin...

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Abstract

The invention relates to a serum-free culture medium for amplification in vitro of a human immune killer cell, wherein the serum-free culture medium is formed by combining and mixing a basal culture medium and a serum replacement, and the serum-free culture medium comprises the basic components of 2 saccharides, 22 amino acids, 6 lipids, 2 nucleosides, 11 vitamins, 16 salts, 1 hormone, 2 protides, hydroxyethylpiperazine ethane sulfonic acid and phenol red. The serum replacement is combined and added into the basal culture medium to prepare the serum-free culture medium. The components of the serum-free culture medium are definite, and the serum-free culture medium supports amplification in vitro of the immune killer cell, achieves high efficiency and stability, promotes the further study and application of biological therapy, promotes the more smooth progressing of the immune cell therapy, and benefits the nation and the people.

Description

technical field [0001] The invention relates to the technical field of medium research and development of modern biotechnology, in particular to a serum-free medium for in vitro expansion of human immune killer cells. [0002] technical background [0003] With the rapid development of modern biotechnology research, biotherapy has become the fourth way to treat tumors after surgery, radiotherapy, and chemotherapy. In the biological treatment of tumors, immune cell therapy is an important part. In recent years, immune cell therapy has shown its important application value in clinic. The immune cell therapy includes tumor infiltrating lymphocytes (TIL), lymphokine-activated killer cells (LAK), cytokine-induced killer cells (CIK), dendritic cells (DC), cytotoxic T lymphocytes ( CTL), T cell receptor gene-modified T cells (TCR-T), and chimeric antigen receptor T cells (CAR-T) cell therapy, among which, TCR-T and CAR-T cell therapy are currently at the forefront adoptive immuno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N5/0646C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/22C12N2500/24C12N2500/25C12N2500/32C12N2500/33C12N2500/34C12N2500/36C12N2500/38C12N2500/40C12N2500/46C12N2500/90
Inventor 蔡海波谭文松陈小东周乐豪
Owner EAST CHINA UNIV OF SCI & TECH
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