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Serum-free medium for in vitro expansion of human immune killer cells

A serum-free medium and cell-killing technology, applied in the field of medium research and development of modern biotechnology, can solve problems such as increasing heterologous infection, achieve the effect of clear components, promote research and application, and promote smoother progress

Active Publication Date: 2021-08-17
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 medium for in vitro expansion of human immune killer cells
  • Serum-free medium for in vitro expansion of human immune killer cells
  • Serum-free medium for in vitro expansion of human immune killer cells

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·hydrochlo...

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-Arginine·HCl L...

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-Arginine·HCl L...

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Abstract

The serum-free medium for the in vitro expansion of human immune killer cells of the present invention is composed of a combination of basal medium and serum substitutes, and its basic components include 2 kinds of sugars, 22 kinds of amino acids, 6 kinds of lipids, 2 A kind of nucleosides, 11 kinds of vitamins, 16 kinds of salts, 1 kind of hormones, 2 kinds of proteins and hydroxyethylpiperazineethanesulfonic acid and phenol red; The serum-free medium of the present invention can be prepared from the basal medium. The invention has clear components, can support the in vitro expansion of immune killing cells, has high efficiency and stability, can promote in-depth research and application of biological therapy, promotes immune cell therapy to proceed more smoothly, and benefits the country 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 Patents(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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