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| Isolation and Primary culture of rat hepatocytes |
| Xiao-Li Liu, Lan-Juan Li and Zhi Chen |
From the Department of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China (Liu XL, Li LJ and Chen Z)
Correspondence: Xiao-Li Liu, MD (Tel: 86-571-87236558) |
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Abstract 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±12×106 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.
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2002, Vol. 1 
