CULTURE AND CHARACTERIZATION OF HUMAN UROTHELIUM IN-VIVO AND IN-VITRO

The aim of this study was to culture human urothelium and generate eno ugh cells for subsequent reconstructive surgery. Using a modification of the Rheinwald-Green method for the routine culture of keratinocytes from patients with burns, we successfully cultured 98% of 57 biopsies from the renal pelvis, ureter, bladder and urethra of paediatric pati ents. The cells could be split one to three up to 9 times at 7-10 day intervals, giving a surface area of 1000 cm(2) after a 2 month culture period. Primary cultures could not be initiated in defined medium MCD B153, although cells initiated using the Rheinwald-Green method could subsequently be propagated in this medium. Cytokeratin patterns in vit ro were similar to those in vivo in the expression of keratins 7, 18 a nd 19 (characteristic of simple epithelia) and keratin 13 (characteris tic of non-cornified stratified epithelia). Cultured urothelium also e xpressed keratin 14 (characteristic of cornified stratified epithelium ) in about 25% of cells and keratin 16 (characteristic of fast-growing cells). These findings indicate that urothelial cells can be propagat ed in vitro for autologous grafting, and the next step is to identify substrates suitable for urothelial cell growth and differentiation and surgical manipulation.