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Macrophage cryopreservation liquid and macrophage cryopreservation method

A macrophage and cryopreservation method technology, applied in the field of cell cryopreservation, can solve the problems of unsuitability for clinical use, cumbersome operation steps, and few passages, and achieve the effects of improving cryopreservation effect, long passage time, and reducing content.

Active Publication Date: 2016-08-31
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using this method to cryopreserve macrophages takes a long time, the operation steps are cumbersome, and more equipment is required; a large amount of bovine serum and a large amount of DMSO are used in the cryopreservation solution, which has great risks and is not suitable for clinical use.
Moreover, the cryopreservation effect is poor, and it often takes a long time for the cells to recover after recovery, and the surface markers are also relatively poor, and the number of passages is small

Method used

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  • Macrophage cryopreservation liquid and macrophage cryopreservation method
  • Macrophage cryopreservation liquid and macrophage cryopreservation method
  • Macrophage cryopreservation liquid and macrophage cryopreservation method

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Effect test

Embodiment 1

[0021] The macrophage cryopreservation solution used in this embodiment contains the following components in the following contents:

[0022] Dextran 2.5mg / mL, Bovine Serum Albumin 10mg / mL, Sorbitol 5v / v%, DMSO 3v / v%, Low Molecular Dextran 10mg / mL, RPMI 1640 92v / v%, Chitosan 3.5mg / mL .

[0023] The method for freezing macrophages using the above-mentioned macrophage freezing solution is as follows:

[0024] Use the macrophage cryopreservation solution of this example to prepare the P1 generation macrophages to a density of 1×10 7 Cells / mL cell suspension, add the cell suspension into a cryopreservation tube, overnight at -80°C, and transfer the cells to a liquid nitrogen tank for storage at -196°C the next day.

[0025] The P1 generation macrophages were prepared by the following method: PBMCs were separated from peripheral blood by Ficoll centrifugation, and RPMI 1640 medium containing 30ng / mL GM-CSF and 10v / v% FBS was used to prepare 1×10 6 Resuspend PBMC at the density o...

Embodiment 2

[0027] The macrophage cryopreservation solution used in this embodiment contains the following components in the following contents:

[0028] Dextran 0.5mg / mL, bovine serum albumin 5mg / mL, sorbitol 2v / v%, DMSO1v / v%, low molecular dextran 5mg / mL, RPMI 1640 97v / v%, chitosan 1mg / mL.

[0029] The method for freezing macrophages using the above-mentioned macrophage freezing solution is as follows:

[0030] Use the macrophage cryopreservation solution of this example to prepare the P1 generation macrophages to a density of 1×10 7 Cells / mL cell suspension, add the cell suspension into a cryopreservation tube, overnight at -80°C, and transfer the cells to a liquid nitrogen tank for storage at -196°C the next day.

Embodiment 3

[0032] The macrophage cryopreservation solution used in this embodiment contains the following components in the following contents:

[0033] Dextran 4mg / mL, bovine serum albumin 20mg / mL, sorbitol 9v / v%, DMSO 4v / v%, low molecular weight dextran 10mg / mL, RPMI 1640 87v / v%, chitosan 6mg / mL.

[0034] The method for freezing macrophages using the above-mentioned macrophage freezing solution is as follows:

[0035] Use the macrophage cryopreservation solution of this example to prepare the P1 generation macrophages to a density of 1×10 7 Cells / mL cell suspension, add the cell suspension into a cryopreservation tube, overnight at -80°C, and transfer the cells to a liquid nitrogen tank for storage at -196°C the next day.

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Abstract

The invention relates to macrophage cryopreservation liquid and a macrophage cryopreservation method. The macrophage cryopreservation liquid is prepared from the following components: glucan, bovine serum albumin, sorbitol, low molecular dextran, chitosan and RPMI 1640. The macrophage cryopreservation method utilizes the macrophage cryopreservation liquid to cryopreserve macrophages. According to the macrophage cryopreservation liquid and the macrophage cryopreservation method, the cryopreservation effect of the macrophages is improved, the survival rate is relatively high after recovery, the growth state of cells is good and the generation time is relatively long; the macrophage cryopreservation liquid does not utilize FBS (Fasting Blood Sugar) so that the safety of clinical transfusion is greatly improved; and the content of DMSO (Dimethylsulfoxide) is reduced and the toxin risks to human bodies are reduced.

Description

technical field [0001] The invention relates to the field of cell cryopreservation, in particular to a macrophage cryopreservation solution and a cryopreservation method. Background technique [0002] Macrophages are white blood cells located in tissues and belong to phagocytic cells, which participate in non-specific defense (innate immunity) and specific defense (cellular immunity) in vertebrates. 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. Therefore, macrophages also have a certain ability to present antigens. [0003] At present, there are few studies on macrophage cryopreservation. In fact, macrophages are extremely sensitive to cryopreservation conditions due to their long digestion time and terminal differentiation. The existing cryopreservation solution for frozen macrophages consists of 90v / v% FBS and 10v / v%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 葛啸虎陈海佳王一飞万桦
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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