Cell cryopreservation method

Abstract

The invention relates to the field of a cell biotechnology, relates to a cell cryopreservation method and particularly relates to an improved rapid freezing method. The cell cryopreservation method can be used for improving the activity of cells under the condition of ensuring that the cell characteristics are unchanged. The cell cryopreservation method has the characteristics that a gauze bag with a hole is adopted, a freezing tube is suspended in a liquid nitrogen steam layer so as to ensure that the cells in the freezing tube are rapidly cooled, and the activity of the resuscitated cells reaches up to 95%-99%; 10% of DMSO (dimethylsulfoxide) and 90% of serum are used as cryopreservation liquid, so that the permeability of a cell membrane to water can be improved, crystals outside the cells are frozen within short time, and the permeation of moisture is prevented from causing the damage to the cells. The cell cryopreservation method is suitable for the long-term storage of all cell lines. Compared with the prior art, the cell cryopreservation method has the advantages that steps are omitted, the technology is simplified, the resuscitated cells have high activity and unchanged cell characteristics, and the long-term storage purpose can be realized.

Description

technical field [0001] The invention relates to the technical field of cell biology, and relates to a cell preservation method, in particular to a cell cryopreservation method. Background technique [0002] Cell cryopreservation is one of the important links in long-term cell culture technology. Studies have shown that using cryopreservation technology to store cells in -196°C liquid nitrogen at low temperature can temporarily remove the cells from the growth state and preserve their cell characteristics. When needed, the cells can be revived and used for experiments. In experimental research, moderately preserving a certain amount of cells can prevent the cells being cultured from being polluted or the cells are lost due to other accidents, which plays a role in cell preservation. At present, the traditional preservation methods commonly used in experimental research involving cell preservation are as follows: first store the cryopreservation tubes with cells in a 4°C envi...

AI Technical Summary

Problems solved by technology

With this method of preservation, the resuscitation activity of cells is 84.15%. Chinese patent CN1944636 discloses a method for cell cryopreservation, wherein: 1. First place the cell cryopreservation tube in an environment of 18-24°C for 20-30 minutes; 2. Then Put it in the liquid nitrogen vapor layer for 8-20 hours, and finally immerse it in liquid nitrogen; Chinese patent CN101070534 discloses that the cell freezing method is 15 minutes at room temperature, put it into the programmed cooling box, store it at -85°C for 4 hours, and then immerse it in the liquid nitrogen. In nitrogen; Chinese patent CN101463340 discloses tissue preservation, liquid nitrogen gasification layer -80 ℃, 5 minutes, into liquid nitrogen, in this patent, the tissue is placed in the gasification layer for too short time, the tissue has not been deep-frozen, and ice crystals are easy to form in the tissue , affect the future recovery of the tissue, reduce cell viability, and cannot be preserved for a long time; Chinese patent CN101711522 discloses tissue preservation, in the liquid nitrogen liquid surface layer for 5-20 minutes, into liquid nitrogen, in this method, cryopreservation tube It is easy to touch liquid nitrogen, and when the cryopreservation tubes for storing cells or tissues are just frozen, as long as they touch a little liquid nitrogen, it is almost impossible for the recovered cells to survive

[0004] The preservation methods of the above-mentioned cells or tissues all adopt the method of gradually cooling down. Practice shows that these preservation methods still have the following defects: the cell viability after recovery of the cells is less than 85%, and the cells cannot be frozen for a long time.