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Serum substitution combination for immune killer cell amplification in vitro

A technology of serum substitute and in vitro expansion, which is applied in the field of serum substitute combination for in vitro expansion of human immune killer cells, can solve the problems of ineffective support for cell growth and proliferation, and achieve high-efficiency support for proliferation, in vitro expansion, Component-specific effects

Active Publication Date: 2016-04-27
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0058] However, studies have found that simply combining the above ingredients cannot effectively support the growth and proliferation of cells

Method used

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  • Serum substitution combination for immune killer cell amplification in vitro
  • Serum substitution combination for immune killer cell amplification in vitro
  • Serum substitution combination for immune killer cell amplification in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] (1) A combination of serum substitutes for the in vitro expansion of immune killer cells, the final concentration of each component is, in milligrams per liter (mg / L):

[0106] Insulin 0.5mg / L;

[0107] Transferrin 5.5mg / L;

[0108] Bovine serum albumin 3000mg / L;

[0109] Ethanolamine 1.22mg / L;

[0110] α-thioglycerol 5.41mg / L;

[0111] Linolenic acid 1mg / L;

[0112] Cholesterol 4mg / L.

[0113] (2) The preparation of the serum substitute combination for in vitro expansion of immune cells can adopt conventional preparation methods, including the following steps:

[0114] 1. According to the content of the components, dissolve insulin in 1% dilute hydrochloric acid solution to make a concentrated solution. The concentration of the concentrated solution is 1000 times the concentration stated in the component. The volume ratio of the medium is 1:100. .

[0115] 2. According to the content of the components, dissolve transferrin, ethanolamine and α-thioglycerol in ul...

Embodiment 2

[0125] (1) A combination of serum substitutes for the in vitro expansion of immune killer cells, the final concentration of each component is, in milligrams per liter (mg / L):

[0126] Insulin 20mg / L;

[0127] Transferrin 5.5mg / L;

[0128] Bovine serum albumin 6000mg / L;

[0129] Ethanolamine 0.61mg / L;

[0130] α-thioglycerol 5.41mg / L;

[0131] Linolenic acid 1mg / L;

[0132] Cholesterol 4mg / L.

[0133] (2) The preparation method of the serum substitute combination for the in vitro expansion of immune killer cells of the present invention (same as Example 1)

[0134] The serum replacement combination prepared in Example 2 is labeled as serum replacement combination B.

[0135] (3) The implementation and comparison of the serum substitute combination for the in vitro expansion of immune killer cells according to the present invention

[0136] 1. Add the above-mentioned serum replacement combination B to the basal medium formed by mixing DMEM / F12 medium and IMDM medium at a vo...

Embodiment 3

[0140] (1) A combination of serum substitutes for the in vitro expansion of immune killer cells, the final concentration of each component is, in milligrams per liter (mg / L):

[0141] Insulin 10mg / L;

[0142] Transferrin 5.5mg / L;

[0143] Bovine serum albumin 6000mg / L;

[0144] Ethanolamine 2.44mg / L;

[0145] α-Thioglycerol 10.82mg / L;

[0146] Linolenic acid 1mg / L;

[0147] Cholesterol 4mg / L.

[0148] (2) The preparation method of the serum substitute combination for the in vitro expansion of immune killer cells of the present invention (same as Example 1)

[0149] The serum replacement combination prepared in Example 3 is marked as serum replacement combination C.

[0150] (3) The implementation and comparison of the serum substitute combination for the in vitro expansion of immune killer cells according to the present invention

[0151] 1. Add the above-mentioned serum replacement combination C to the basal medium formed by mixing DMEM / F12 medium and IMDM medium at a ...

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Abstract

The invention provides a serum substitution combination for immune killer cell amplification in vitro. Insulin with the final concentration being 0.5-20 mg / L, transferrin with the final concentration being 0.69-22 mg / L, bovine serum albumin with the final concentration being 3000-6000 mg / L, ethanolamine with the final concentration being 0.61-2.44 mg / L, alpha-thioglycerol with the final concentration being 5.41-10.82 mg / L, linolenic acid with the final concentration being 0.5-5.0 mg / L and cholesterol with the final concentration being 0.8-20 mg / L are added into a conventional culture medium. In the serum substitution combination, ferric ammonium citrate with the concentration being 0.46-2.3 mg / L, beta-disodium glycerophosphate with the concentration being 300-1500 mg / L and putrescine with the concentration being 0.25-1.25 mg / L can be added. The components are definite, can be added into different culture media such as an RPMI1640 culture medium, a DMEM / F12 culture medium and an IMDM culture medium, can support immune killer cell amplification in vitro, and are efficient and universal. In vitro serum-free amplification of immune killer cells can be achieved, the biological therapy of tumors is promoted, and conduction of the somatic cell therapy is promoted particularly.

Description

technical field [0001] The invention relates to the technical field of culture medium research and development of modern biotechnology, in particular to a serum substitute combination for in vitro expansion of human immune killer cells. technical background [0002] With the development of biotherapy tumor technology, biotherapy has become the fourth treatment for tumors after surgery, radiotherapy, and chemotherapy. Immune cell therapy is an important part of the biological treatment of tumors. In recent years, immune cell therapy has shown important clinical application value. [0003] The immune cell therapy includes tumor infiltrating lymphocytes (TIL), lymphokine-activated killer cells (LAK), cytokine-induced killer cells (CIK), dendritic cells (DC) and cytotoxic T lymphocytes (CTL) )Treatment. Among them, tumor adoptive immunotherapy based on autologous cytokine-induced killer cells (CIK) has gradually become one of the most active applications and research projects ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0037C12N5/0646C12N2500/90
Inventor 蔡海波陈小东谭文松张伟伟施镇
Owner EAST CHINA UNIV OF SCI & TECH
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