Frozen stock solution for nerve cells and freezing storage method

Abstract

The invention discloses cell frozen stock solution for freezing and storing nerve cells and a freezing storage method, wherein, the cell frozen stock solution and the freezing storage method are used for freezing and storing the nerve cells, especially primary animal nerve cells that are isolated freshly, such as hippocampus nerve cells. The frozen stock solution is formed by adding basic medium,fetal bovine serum, dimethyl sulfoxide,6-fructose phosphate and flavonoid compounds to the basic medium; 6-fructose phosphate (w/w) achieves the function of improving cell viability (effective cell rate) when the frozen cells are recovered; and in addition, the flavonoid compounds achieve definite oxidation resistance and can prevent free radical from damaging the cells. The cell frozen stock solution and the freezing storage method can improve the survival rate and the cell function of the nerve cells through freezing storage, the survival rate of unfreezing cells can be increased to reach more than 95 percent, the frozen stock solution for the nerve cells can preserve the nerve cells for a long period, the biological activity of the nerve cells can be ensured, and very high utility values can be achieved.

Description

technical field [0001] The invention relates to a cryopreservation solution and a cryopreservation method for cells. Background technique [0002] As an effective method to preserve cells, cell freezing technology has been widely used in the field of biology. The low-temperature cryopreservation technique is to add dimethyl sulfoxide and serum to the cell suspension, put it into a cryopreservation tube, put it in a -80°C refrigerator overnight, and then transfer it to liquid nitrogen for storage. For many kinds of cells, the cells can still survive through the freeze-thaw process and can continue to be cultured by adding suitable freezing liquid. However, in most cases, it is suitable for cells that can be cultured in vitro and survive for a long time, such as myeloma cells and hybridoma cells. Chinese Patent Announcement No. CN 1164739C discloses a cell cryopreservation solution for improving the recovery rate of frozen cells. sulfone. The cells targeted are common anim...

AI Technical Summary

Problems solved by technology

Glucose is an energy substance necessary for the energy metabolism of neurons. Glucose is beneficial to the growth of neurons, but if the concentration of glucose is too high, it will easily increase the chance of pollution

The survival rate of nerve cells after cryopreservation and resuscitation is poor, and the cell activity is low. At present, an ideal nerve cell cryopreservation medium and cryopreservation method has not been explored. The reasons are mainly related to the toxicity of the cryopreservation solution and the lack of cell energy.

[0004] Chinese patent CN1966674A discloses a method for cryopreserving neural stem cells. The invention adopts 15% 2×DMSO into collagen medium to freeze-dry. Dimethyl sulfoxide (DMSO) is a commonly used protective agent for cell cryopreservation. It is a colorless transparent liquid with strong polarity, which can significantly change the activities of various enzymes in the cells, thereby facilitating the cryopreservation of cells. However, if the concentration of dimethyl sulfoxide is too high (≥10%), it is harmful The stored cells have severe toxic effects and interact with protein hydrophobic groups, leading to protein denaturation

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