In-vitro cultivation of urothelial cells for genito-urinary tract reconstructive surgery: an experimental study

ABSTRACT

An in-vitro system for the cultivation of urothelial cells has been developed. Urothelial cells were isolated from 30 tissue samples obtained from the urinary bladders of six adult New Zealand rabbits that underwent partial cystectomy. The cells from 15 samples were grown in M-199 culture media and that from the remaining 15 samples were grown over human placental membranes measuring 3 x 4 x 0.1 centimeter (cm) contained in the same media. Confluent cell monolayers covering the entire base of the plastic multiwells and the entire amniotic membranes were produced within 7 days respectively in all cases. Subsequent subcultures likewise achieved the same confluency within another 7 days. Morphological analysis of all cultures by inverted phase contrast microscopy revealed cells of epithelioid nature. Cells from the media were likewise concentrated by centrifugation and smeared over glass slides, heat-dried and fixed with absolute ethanol. The smears underwent Giemsa staining and were examined under light microscopy and confirmed the presence of urothelial cells. The cultured urothelial graft developed from this study is a potentially useful material for future in-vivo researches involving genito-urinary tract reconstructive surgeries.