Isolation of oval cells by centrifugal elutriation and comparison with other cell types purified from normal and preneoplastic livers.
Oval cells and biliary epithelial cells were isolated from livers of rats fed a choline-deficient diet containing 0.1% ethionine and from normal rat livers, respectively. Nonparenchymal cell suspensions prepared from these livers by collagenase perfusion followed by digestion of undissociated tissue with 0.1% collagenase, 0.1% Pronase, and 0.004% DNase I were separated into six fractions by centrifugal elutriation. Cells in each fraction were characterized histochemically for gamma-glutamyl transpeptidase, peroxidase, alkaline phosphatase, and glucose-6-phosphatase activities, and for albumin and alpha-fetoprotein by immunocytochemical methods. Cells from Fraction 5 of the elutriation procedure had various features predicted for oval cells and were selected for further studies. The cell yield in this fraction, from each preneoplastic liver, was 5.7 X 10(7) cells, 93 +/- 2% of which were gamma-glutamyl transpeptidase positive, 6 +/- 1% peroxidase positive, 61% albumin positive, and 29% alpha-fetoprotein positive. Cells in this fraction have a median diameter of 13.1 micron and are diploid and cycling. The majority of these cells has morphological features characteristic of biliary epithelial cells, although some cells display features intermediate between duct cells and hepatocytes. Nucleic acid hybridization using specific probes revealed that these cells contain albumin and alpha-fetoprotein messenger RNAs, while hepatocytes from normal and preneoplastic liver contain only albumin messenger RNA. Biliary cells obtained from normal livers do not contain albumin messenger RNA. The large-scale purification and characterization of cell populations from preneoplastic livers is an important step in elucidating the cellular derivation of liver tumors.
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002796:Choline Deficiency / Q000473:pathology; D004847:Epithelial Cells; D005001:Ethionine; D005704:Gallbladder / Q000166:cytology; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008113:Liver Neoplasms / Q000473:pathology*; D008297:Male; D008854:Microscopy, Electron; D011003:Ploidies; D011230:Precancerous Conditions / Q000473:pathology*; D051381:Rats; D011919:Rats, Inbred Strains
D005001:Ethionine