Evaluation of viability and proliferative activity of human urothelial cells cultured onto xenogenic tissue-engineered extracellular matrices.
OBJECTIVE
To evaluate the viability and proliferative activity of human urothelial cells (HUCs) cultured on tissue-engineered extracellular matrix scaffolds and to assess the potential of extracellular matrixes to support the growth of HUCs in their expected in vivo urine environment.


METHODS
HUCs were obtained by bladder biopsy and cultured onto the luminal and abluminal surfaces of decellularized porcine small intestinal submucosa (SIS) and porcine urinary bladder matrix (UBM). In addition, HUCs were cultured in optimal in vitro growth conditions and in their expected in vivo urine environment. The attachment, viability, and proliferative activity of HUCs were evaluated and compared using quantitative viability indicators and fluorescent markers for intracellular esterase activity and plasma membrane integrity.


RESULTS
The luminal and abluminal surfaces of the UBM demonstrated significantly greater HUC viability and proliferative activity compared with the luminal and abluminal surfaces of the SIS grafts (P < .0001). Culture of HUCs in a simulated in vivo urine environment significantly affected cell viability (P < .0001). Proliferative activity was immeasurable on cell-seeded scaffolds that were cultured in a urine environment after 48 hours of growth (P < .0001).


CONCLUSIONS
This is the first comparative report of UBM and SIS. Our results have demonstrated that UBM has significantly greater regenerative potential for HUCs compared with SIS. However, the perceived potential for extracellular matrixes in reconstructive urology might be limited by their inability to induce urothelial regeneration in a urine environment.
Department of Urology, Mid-Western Regional Hospital, Limerick, Ireland.
Niall F
NF
Davis
Anthony
A
Callanan
Barry B
BB
McGuire
Hugh D
HD
Flood
Tim M
TM
McGloughlin
D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004950:Esterases / Q000378:metabolism; D004996:Ethidium / Q000031:analogs & derivatives / Q000378:metabolism; D005109:Extracellular Matrix; D006801:Humans; D007150:Immunohistochemistry; D007364:Intercalating Agents / Q000378:metabolism; D007413:Intestinal Mucosa; D009130:Muscle, Smooth / Q000166:cytology; D010078:Oxazines; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014183:Transplantation, Heterologous; D019459:Urothelium / Q000166:cytology*; D014966:Xanthenes
D007364:Intercalating Agents; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; C018533:ethidium homodimer; D004950:Esterases; D004996:Ethidium