Culture medium for culturing hepatocytes and preparation method thereof

Abstract

The invention relates to the technical field of a cell culture medium and particularly relates to a culture medium for culturing hepatocytes and a preparation method thereof. The culture medium comprises a basic culture medium and additives in the basic culture medium. The additives comprise, on the basis of final concentration, 2 to 10 milligrams per liter of cholesterol, 30 to 100 micrograms per liter of lipoic acid, 0.2 to 0.5 milligrams per liter of chondroitin sulfate, 13 to 16 micromoles per liter of antibiotic, 0.9 to 1.3 milligrams per liter of insulin, 10 to 17 milligrams per liter of catalase, 2 to 7 micrograms per liter of all-transretinoic acid, 14 to 22 micrograms per liter of fibronectin, 180 to 420 milligrams per liter of fat-type bovine serum protein, 10 to 60 micrograms per liter of 2-mercaptoethanol, 3 to 12 milligrams per liter of rapamycin, 3 to 9 milligrams per liter of folic acid, 0.2 to 1.6 milligrams per liter of progesterone, 5 to 23 milligrams per liter of trace elements and 20 to 35 milligrams per liter of a cell growth factor. In the culture medium, stem cells can well grow and cell form and density are the same to those in a serum culture medium. The culture medium solves the problems of the traditional serum-containing culture medium. The culture medium has a reasonable formula. The components produce synergism, provide enough nutrients and a good environment for cell grow and proliferation and improve viability of hepatocytes.

Description

technical field [0001] The invention relates to the technical field of cell culture medium, in particular to a culture medium for culturing hepatocytes and a preparation method thereof. Background technique [0002] Using animal cell culture technology to verify the most efficient and feasible method of compound toxicology experimental testing so far. With the continuous expansion of the application field and market demand of biotechnology in the toxicological safety evaluation of compounds, improving animal cell culture and toxicology tests and extending the maintenance time to obtain reliable results have become the core of optimizing animal cell culture processes and toxicology experiments. [0003] As we all know, the economy and efficiency of animal cell culture and toxicology test are determined by the medium, and the optimization of the medium is the core link in the establishment of efficient animal cell culture and toxicology test process. Therefore, it is an impor...

AI Technical Summary

Problems solved by technology

Chinese patent application CN105087465A discloses a hepatocyte serum-free medium, which includes the following components: basal medium 500mL; sericin 0.05-0.5%; dexamethasone 0.1-1000nmol / mL; hepatocyte growth factor 5-20ng / mL; epidermal growth factor 10 ~ 50ng / mL; penicillin and streptomycin 100U / mL, these serum-free media are used for hepatocyte culture, but they are expensive, which is not conducive to daily application, widespread promotion and hepatocyte industry culture and other issues

Therefore, when performing animal cell culture and toxicology experiments, it is difficult for most serum-free media to provide cells with the nutrients they need in a sufficient and balanced manner.

Medium utilization rate is very low, resulting in great waste