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Macrophage induced culture medium and macrophage culture method

A technology for inducing culture medium and macrophages, which can be used in animal cells, vertebrate cells, blood/immune system cells, etc. It can solve the problems of low purity of macrophages and insignificant cell expansion, and achieve strong antigen presentation. Ability and killing ability, strong proliferation ability, high purity effect

Inactive Publication Date: 2016-03-02
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the purity of the macrophages cultured in the prior art is very low, and the cell expansion is not obvious

Method used

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  • Macrophage induced culture medium and macrophage culture method
  • Macrophage induced culture medium and macrophage culture method
  • Macrophage induced culture medium and macrophage culture method

Examples

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Embodiment 1

[0028] Embodiment 1, human peripheral blood macrophage induction medium and culture method

[0029] A medium for inducing macrophages, comprising the following components in the following contents: 500 U / mL GM-CSF, 5% plasma by volume, and X-VIVO15 serum-free medium as the balance.

[0030] CD14 isolated from PBMC using the above macrophage induction medium + A method for inducing cells to be macrophages, comprising the steps of:

[0031] S1. CD14 was sorted out from PBMC with a CD14 sorting kit (purchased from STEMCELL) + Cells; resuspend CD14 in X-VIVO15 serum-free medium + cells, by 5×10 5 The density of cells / mL was inoculated into T25 culture flasks, using macrophage induction medium at 37°C, 5% CO 2 Cultivate under certain conditions, observe regularly, and perform cell rehydration every 2-3 days.

[0032] S2. After culturing until the 5th day, add the K562 cell lysate supernatant for stimulation, preferably the final protein concentration in the K562 cell lysate su...

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Abstract

The invention discloses a macrophage induced culture medium and a macrophage culture method. The macrophage induced culture medium contains a GM-CSF (granulocyte-macrophage colony stimulating factor), plasma and a serum-free basal culture medium. The macrophage culture method includes using tumor cell lysate supernatant for stimulating macrophages. The macrophage induced culture medium and the macrophage culture method have the advantages that a specific concentration of GM-CSF and the plasma are used for inducing CD14<+>cells and the tumor cell lysate supernatant is used for stimulating the CD14<+>cells, so that macrophage proliferation is promoted, and macrophage purity is also improved.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a macrophage induction medium and a culture method. Background technique [0002] Macrophages are tissue-resident white blood cells derived from monocytes. Macrophages are one of the three major antigen-presenting cells, which have a wide range of biological effects, such as anti-infection, anti-tumor, participation in immune response and immune regulation, and are important cells involved in non-specific and specific immunity. The main function of macrophages is to phagocytize (i.e. phagocytize and digest) cell debris and pathogens in the form of fixed cells or free cells, and activate lymphocytes or other immune cells to make them respond to pathogens. Macrophages are immune cells with multiple functions. They are easy to obtain, easy to culture, and can be purified. They are important objects for the study of cell phagocytosis, cellular immunity, and molecular immunology. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786
Inventor 王一飞陈海佳葛啸虎罗二梅
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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