Isolation and primary culture of rat hepatocytes.
OBJECTIVE
To find out an optimal condition for isolation and primary culture of hepatocytes.


METHODS
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.


CONCLUSIONS
High yield of hepatocytes can be isolated with two-step collagenese perfusion. Hepatozyme-SFM is suitable for sustained growth of hepatocytes.
Liu
Xiao-Li
XL
Department of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Li
Lan-Juan
LJ
Chen
Zhi
Z
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D017364:Collagenases; D022781:Hepatocytes / Q000166:cytology*; D019164:Liver, Artificial; D051381:Rats; D017207:Rats, Sprague-Dawley
D017364:Collagenases