Culture method for long-term maintenance of functional state of human primary hepatocytes
A technology of primary hepatocytes and functional status, which is applied in the field of medicine and can solve problems such as complex operation techniques of culture equipment
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[0024] 1. Experimental materials
[0025] The recombinant protein HGF used in this example was purchased from Peprotech; the remaining small molecule compounds such as Y27632, A83-01 and CHIR99021 were purchased from TargetMol; the primary cells used were purchased from BioreclamationIVT, and specific cell culture basal (i.e. hepatocyte expansion medium) culture; penicillin and streptomycin double antibiotics and the basal medium used for subsequent primary hepatocyte maintenance experiments were purchased from Yuanpei Biotechnology Company; fetal bovine serum (FBS) was purchased from in Gibco; Matrigel used in cultured primary hepatocytes was purchased from BD.
[0026] 2. Experimental method
[0027] 1. Cell recovery and inoculation
[0028] After the frozen cells were taken out of the liquid nitrogen tank, they were quickly shaken in a water bath at 37°C until they were completely melted, centrifuged at 700rpm at 4°C for 3 minutes, the supernatant was discarded, and 1ml o...
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