title: [Establishment and growth regulation of rat hepatocyte colony in vitro].
abstract: OBJECTIVE
  To investigate the conditions essential for culturing hepatocytes colony in vitro, and to study the growth regulation mechanisms of hepatocyte colony.
  
  
  METHODS
  Rat hepatocytes were isolated by two-step in situ preperfusion and collagenase circulatory perfusion. The effects of hepatopoietin (HPN), nicotinamide (NA) and dimethyl sulfoxide (DMSO) on DNA synthesis, mitotic activity, morphology (under inverted microscope) and utrastructure (under TEM) of the hepatocytes were investigated in chemically defined culture medium.
  
  
  RESULTS
  The time course of DNA synthesis in cultured hepatocytes showed that 3H-TdR incorporation was dramatically enhanced by 10 mmol/L NA, presenting 2 peaks at 60 and 84 h respectively. The plateau of DNA synthesis was diminished in the presence of DMSO, but a peak occurred again at 132 h upon NA treatment. After cell culture in the presence of HPN, NA, and DMSO for 72 h, the hepatocytes presented sustained regular bipolar mitosis, with considerable mitotic activity at 168 h. The growth characteristics of hepatocyte colony, in addition to its potential for expansion, were captured by both light and electron microscopy on day 28 of cell culture.
  
  
  CONCLUSIONS
  The reciprocal actions of NA and DMSO can control the proliferation of HPN-stimulated hepatocytes, which can be used for studying human hepatocyte metabolism, cytotoxicity, biotransformation and mutagenesis, and may provide experimental evidences for the treatment of liver failure and genetic liver diseases with in vitro hepatocyte clones.
authors.affiliation: Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China. tangxf@fimmu.com
authors.forename: Xi-feng
authors.initials: XF
authors.lastname: Tang
authors.forename: Dong-yao
authors.initials: DY
authors.lastname: Zhou
authors.forename: Zhi-min
authors.initials: ZM
authors.lastname: Xu
authors.forename: Xiao-dong
authors.initials: XD
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002455:Cell Division / Q000502:physiology; D004247:DNA / Q000096:biosynthesis; D017228:Hepatocyte Growth Factor / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D051381:Rats
chemical_list: D017228:Hepatocyte Growth Factor; D004247:DNA