05 January 2022 1 6K Report

I am isolating primary hepatocytes from mice. I am using the two-step collagenase perfusion method and after seeding the cells in the 12-well plate, I am seeing two types of cell morphologies, one with hepatocyte morphology and another with fibroblast-like morphology. Since according to my knowledge it's not possible to passage primary hepatocytes, it is difficult to get a pure culture after passaging. The alternative to this problem is that I sort the hepatocytes before seeding. But I couldn't find any marker specific for mice primary hepatocytes. We can't use the regular adherent markers such as EPCAM and all because it is present in fibroblasts also. So any advice or suggestion will be highly appreciated. Thank you in advance.

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