title: Isolation and primary culture of rat hepatocytes.
abstract: 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.
authors: Department of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
authors: Xiao-Li
authors: XL
authors: Liu
authors: Lan-Juan
authors: LJ
authors: Li
authors: Zhi
authors: Z
authors: Chen
mesh_terms: 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
chemical_list: D017364:Collagenases