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Updated: Aug 30, 2026
Isolation and Primary Culture of Rat Hepatic Cells
Published on: June 29, 2012
Isolation and primary culture of rat hepatocytes
Xiao-Li Liu1, Lan-Juan Li, Zhi Chen
1Department of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Objective:
To find out an optimal condition for isolation and primary culture of hepatocytes.
Method:
Rat hepatocytes were isolated by a two-step collagenase perfusion, and cultured in hepatozyme-SFM. The reduction of MTT to formazan salt was examined. Supernatant medium was collected for analysis of alanine amino transferase (ALT) and ureagenesis.
Results:
The two-step collagenase perfusion yielded 39+/-12x10(6) cells/g liver tissue with a viability of 88%+/-2%. Fine morphology and stable urea synthesis for one week could be achieved when hepatocytes were cultured in hepatozyme-SFM.
Conclusion:
High yield of hepatocytes can be isolated with two-step collagenese perfusion. Hepatozyme-SFM is suitable for sustained growth of hepatocytes.
