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Macrophage medium and culture method

A technology of macrophage and culture method, which is applied in the field of cell culture, can solve the problems of limited macrophage culture expansion, unsuitability for large-scale culture, labor and time consumption, etc., and achieves shortened culture time and operation time, and is easy to operate , the effect of reducing waste

Inactive Publication Date: 2016-07-20
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This prior art solution is limited to macrophage culture and expansion, and is a method for small-scale culture, which is not suitable for large-scale culture. When the demand for cells increases, the operation is cumbersome and consumes a lot of manpower and time.

Method used

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  • Macrophage medium and culture method
  • Macrophage medium and culture method
  • Macrophage medium and culture method

Examples

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Effect test

Embodiment 1

[0026] A macrophage culture medium, containing the following components of the following content: DMEM medium 90v / v%, fetal bovine serum 10v / v%, GM-CSF30ng / mL, lipopolysaccharide 100μg / L, IL-1420ng / mL.

[0027] A method for culturing macrophages, using the above-mentioned macrophage culture medium, microcarriers and rocking bag cell bioreactors, specifically comprising the following steps:

[0028] S1. Use lymphocyte separation medium to separate PBMC from peripheral blood; use the above-mentioned macrophage culture medium to resuspend PBMC so that the density of PBMC is 0.5×10 6 -2×10 6 cells / mL, inoculate it in a T75 culture flask, culture for 7 days, change the medium every 3-4 days; discard the supernatant on the 7th day, add trypsin-EDTA digestion solution to digest until the cells are completely detached, add 3 times the volume The FBS terminated the digestion, and the supernatant was removed by centrifugation; the mass concentration percentage of EDTA in the trypsin-...

Embodiment 2

[0032] A macrophage culture medium, containing the following components of the following content: DMEM medium 75v / v%, fetal calf serum 25v / v%, GM-CSF15ng / mL, lipopolysaccharide 80μg / L, IL-1410ng / mL.

[0033] A method for culturing macrophages, using the above-mentioned macrophage culture medium, microcarriers and rocking bag cell bioreactors, specifically comprising the following steps:

[0034] S1. Use lymphocyte separation medium to separate PBMC from peripheral blood; use the above-mentioned macrophage culture medium to resuspend PBMC so that the density of PBMC is 0.5×10 6 -2×10 6 cells / mL, inoculate it in a T75 culture flask, culture for 7 days, change the medium every 3-4 days; discard the supernatant on the 7th day, add trypsin-EDTA digestion solution to digest until the cells are completely detached, add 3 times the volume The FBS terminated the digestion, and the supernatant was removed by centrifugation; the mass concentration percentage of EDTA in the trypsin-EDT...

Embodiment 3

[0038] A macrophage culture medium, containing the following components of the following content: DMEM medium 95v / v%, fetal bovine serum 5v / v%, GM-CSF50ng / mL, lipopolysaccharide 160μg / L, IL-1450ng / mL.

[0039] A method for culturing macrophages, using the above-mentioned macrophage culture medium, microcarriers and rocking bag cell bioreactors, specifically comprising the following steps:

[0040] S1. Use lymphocyte separation medium to separate PBMC from peripheral blood; use the above-mentioned macrophage culture medium to resuspend PBMC so that the density of PBMC is 0.5×10 6 -2×10 6cells / mL, inoculate it in a T75 culture flask, culture for 7 days, and change the medium every 3-4 days; discard the supernatant on the 7th day, add trypsin-EDTA digestion solution to digest until the cells are completely detached, add 3 times volume of FBS to terminate the digestion, and centrifuge to remove the supernatant; the mass concentration percentage of EDTA in the trypsin-EDTA diges...

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Abstract

The invention relates to a macrophage medium and a culture method. The macrophage medium contains a DMEM medium, FBS, GM-CSF, lipopolysaccharide and interleukin-14. In the culture method of macrophage, the macrophage is cultured by use of the macrophage medium provided by the invention, a microcarrier and a shaking bag type cell culture bioreactor. By adopting the macrophage medium and culture method provided by the invention, the culture time is shortened, the consumption of human resources is reduced, and large-scale culture of macrophage can be realized.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a macrophage culture medium and a culture method. Background technique [0002] Macrophages are one of the three major antigen-presenting cells and are important cells involved in nonspecific and specific immunity. Their main function is to phagocytize (i.e. phagocytize and digest) cell debris and pathogens in the form of fixed or free cells, and activate lymphocytes or other immune cells to respond to pathogens. Because macrophages have strong phagocytosis and antigen presentation functions, they have great research value in immunology. [0003] Due to the limitation of culture conditions, it is very difficult to obtain a large number of macrophages, and there is a lack of methods for large-scale culture of macrophages, which limits the application of macrophages. The method for cultivating macrophages in the prior art is as follows: isolate mononuclear cells (PBMCs) in peripheral ...

Claims

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Application Information

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IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2501/22C12N2501/2314C12N2501/90C12N2531/00
Inventor 葛啸虎陈海佳王一飞万桦
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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