Cryopreservation solution and cryopreservation resuscitation method for liver primary cells

Abstract

The invention discloses a cryopreservation solution and cryopreservation resuscitation method for liver primary cells and relates to the field of biotechnologies. The cryopreservation solution is prepared according to the following formula, including 1,000g of dH2O, 15.6g of DMEM / F12 culture medium powder, 1.2g of sodium bicarbonate, 300ng of dexamethasone, 10 micrograms of liver primary cell growth factor, 25mg of insulin, 0.3g of glutamine, 25mg of gentamycin, 50mg of penicillin, 25mg of clarithromycin, 0.1g of DMSO, 10g of PEG6000, 1g of polyvinyl pyrrolidone and 1g of 1,3-propylene glycol. According to the cryopreservation solution and cryopreservation resuscitation method for the liver primary cells, the problem that the liver primary cells are relatively low in survival rate after cryopreservation resuscitation and cannot adhere to walls for drug metabolism study is solved, and the developed new cell cryopreservation solution formula and the corresponding resuscitation method can enable the cryopreservation resuscitation survival rate of the cells to be over 80%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cryopreservation solution of liver primary cells and a cryopreservation recovery method thereof. Background technique [0002] Drug metabolism is mainly completed by phase I and phase II drug metabolizing enzymes in the liver, and CYP450 enzymes play an important role in the biotransformation process, and more than 90% of drug oxidation is catalyzed by CYP450 enzymes, which are the most stable in the body. Important metabolic enzymes, primary liver cells retain all CYP450 enzymes, and are the best in vitro model for studying drug metabolism. Using primary liver cells in vitro to study drug metabolism pathways and clearance rates has a relationship with liver metabolism under physiological conditions in vivo. comparability. The data obtained by multiple laboratories from different species (including human) in vitro primary liver cell models show that the species differe...

AI Technical Summary

Problems solved by technology

The traditional cell cryopreservation solution is composed of DMEM or RPMI-1640 adding a certain proportion of dimethyl sulfoxide and fetal bovine serum, and then the cells are frozen by the programmed cooling method. Primary cells have a low survival rate and cannot adhere to the wall for drug metabolism research, which needs further improvement

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