Abstract:

BACKGROUND: Methods to separate the transitional epithelial cells include enzyme digestion method, tissue block method and spinning scraping method.
OBJECTIVE: To investigate the best approach to culture and passage the human bladder transitional cells in vitro.
METHODS: Human bladder transitional epithelial cells were cultured in MEM-F12 and KSFM media. Then the cell morphological changes, growth and proliferation were observed. The bladder transitional epithelial cells were identified by immunofluorescent staining based on the presence of uothelium specific cell markers AE1 and Uroplakin III, the latter one specifically expressed in urothelial tissue. The cells were subcultured by 4-step trypsin digestion method and tissue block replantation method, respectively.
RESULTS AND CONCLUSION: The cells in EME-F12 media grew well, and the cell climbing-out and cell fusion occurred earlier than cells in KSFM media. The cells were identified as transitional epithelial cells by immunofluorescence. Passage cells subcultured by trypsin digestion began to adhere to the wall at 18-24 hours after passage. The cells stopped growing after adherence. Cellapoptosis appeared after 60-72 hours. The passage cells subcultured by tissue block replantation displayed a stable and fast growth. The cell began to climb out at 24-48 hours. Cell fusion rate achieved 80% on 10-16 days. Cells began to degenerate at the fourth passage. These results demonstrate that tissue block cultivation and replantation based on DMEM-F12 is an economic and effective way to culture and subculture the human bladder transitional cells.