title: Isolation and cultivation of adult primary bovine hepatocytes from abattoir derived liver.
abstract: The aim was to establish a cell culture of adult primary bovine hepatocytes obtained from liver following slaughter and to cultivate the cells in a sandwich culture. Cells and medium samples were taken after separation of cells (day 0), during monolayer (days 1, 2 and 3) and during sandwich culture (days 1, 2, 3, 4, 7, 10 and 14). The mRNA expression ofBAX,BCL2L,FAS,IGF-1andGHR1Awas measured as well as urea and LDH. Hepatocytes were obtained by using a two-step collagenase perfusion and were purified thereafter by density gradient centrifugation. The viability was 68.2 ± 9.5 %. In sandwich culture, cells have a typical polygonal hepatocyte-like shape, build cell-cell contacts, and show irregularity of cell borders suggesting bile canaliculi generation. TheBAXmRNA expression increased on day 1 as well but decreased steadily until day 3 and remained constant for 14 days. Urea- and LDH-concentrations increased from day 4 to day 7. In conclusion, we found that it is possible to gather viable primary hepatocytes from adult bovine liver after slaughter, and that cells gathered this way show typical morphologies, urea-production and low LDH-leakage especially at day 4 in a sandwich system.
authors.affiliation: University of Veterinary Medicine, Clinic for Cattle, Endocrinology, Bischofsholer Damm 15, 30173 Hannover, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Ehrhardt
authors.affiliation: University of Veterinary Medicine, Clinic for Cattle, Endocrinology, Bischofsholer Damm 15, 30173 Hannover, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Schmicke
keywords: cattle; cell culture; hepatocyte; liver; sandwich