Anchorage-dependent cell cryoprotectant, cryopreservation method and thawing method

Abstract

The invention discloses anchorage-dependent cell cryoprotectant. The anchorage-dependent cell cryoprotectant is prepared from DMS0, 1,2-propanediol, polyethylene glycol, BSA and DMEM culture medium. The invention also discloses a cryopreservation method of anchorage-dependent cells. The method comprises the following steps: adding the cryoprotectant into the anchorage-dependent cells, uniformly mixing, and cryopreserving. The invention also discloses a thawing method of the anchorage-dependent cells. The thawing method comprises the following steps: (1) collecting the cryopreserved anchorage-dependent cells, performing water bath, heating, and thawing; and (2) removing the cryoprotectant in the anchorage-dependent cell in step (1), cleaning by using a PBS solution, removing the cryoprotectant remaining in the anchorage-dependent cell, thus obtaining the thawed anchorage-dependent cell. After the anchorage-dependent cells are cryopreserved by adopting the cryoprotectant of the invention, the anchorage-dependent cells can be stored at a low temperature for a long time and can be fetched for use according to the requirement, and the anchorage-dependent cells can be used by virtue of asimple recovery program (less than or equal to 10 minutes); and after being cryopreserved, the anchorage-dependent cells can be directly transported at the low temperature, so that the convenient immediately-available cells can be provided for the laboratory with no cell culture condition.

Description

technical field [0001] The invention relates to the technical field of cell preservation, in particular to a cryopreservation solution, a cryopreservation method and a thawing method for adherent cells. Background technique [0002] During the culture process, cells are divided into two types: adherent growth and suspension growth according to the growth mode, and most of the cells are adherent growth. The traditional freezing method for adherent cultured cells is generally to digest the adherent cells first, then collect the cell digestion solution and centrifuge, then remove the supernatant and add freezing liquid, completely wrap the cells in the freezing liquid, and put them into a cryopreservation tube The temperature was gradually lowered, and finally put into liquid nitrogen for long-term storage. This method is very successful for the long-term preservation of germplasm resources. The preservation time can be as long as ten or even twenty years, and the cells are st...

AI Technical Summary

Problems solved by technology

[0005] The method of digesting the cells, suspending them in the medium and then freezing them can only be used to preserve the cell lines. The cells need to be resuscitated before each use, which cannot meet the requirements of taking them at any time.

[0006] Viruses are strictly intracellular parasites, so in the clinical virus isolation and identification system, a large number of cells are needed for virus culture, but the cells also have a certain life span, in order to avoid the occurrence of cell growth and morphology caused by long-term in vitro culture of cells Degeneration or transformation, requiring frequent, tedious, and time-consuming passaging of cells

In addition, in clinical work, it is necessary to accurately predict the amount of clinical specimens, but there are often situations where the quality of cells between batches is unstable, the supply of cells is insufficient, or the preparation of cells is too much

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