Growth and characterization of porcine urinary bladder epithelial cells in vitro.
Dividing long-term monolayer cultures of porcine urinary bladder epithelial cells were obtained by a combined mechanical and enzymatic isolation method. The serum-free cultured cells were investigated morphologically and characterized according to their growth characteristics and enzymatic functions. Investigations over a period up to 12 wk demonstrated that the cells regain their in vivo polarization with apically situated membrane vesicles and tight junctions between neighboring cells when they have built up a confluent monolayer. Activities of most marker enzymes for cell-differentiated status and function of such cells observed over a period of 4 wk in culture were conserved compared with the original tissue. Lactate dehydrogenase activity release into the medium was at low levels (< or = 5% of the total amount), indicating a good membrane integrity and cell viability. The chromosome set (2n = 38) did not change significantly during the first 5 wk, but, with additional culture time, the degree of polyploid and polynucleated cells increased comparable to the in vivo situation, in which the apical cell layer of the bladder mucosa also showed a high degree of polynucleation and polyploidy, indicative of a senescence process.
Guhe
C
C
Institut für Arbeitsphysiologie, Universität Dortmund, Germany.
Föllmann
W
W
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D015925:Cryopreservation; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D003896:Desmosomes / Q000648:ultrastructure; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D007621:Karyotyping; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008677:Metaphase; D008854:Microscopy, Electron; D009245:NADH Dehydrogenase / Q000378:metabolism; D015388:Organelles / Q000648:ultrastructure; D013552:Swine; D013997:Time Factors; D001743:Urinary Bladder / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure
D003470:Culture Media; D007770:L-Lactate Dehydrogenase; D009245:NADH Dehydrogenase