Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle-urothelial cell interactions.
The objective of this study was to evaluate the ability of bladder acellular matrix (BAM) to support the individual and combined growth of primary porcine bladder smooth muscle (SMC) and urothelial (UEC) cells. An in vitro co-culture system was devised to evaluate the effect of UEC on (i) SMC-mediated contraction of BAM discs, and (ii) SMC invasiveness into BAM. Cells were seeded onto BAM discs under 4 different culture conditions. Constructs were incubated for 1, 7, 14 and 28 days. Samples were then harvested for evaluation of matrix contraction. Immunohistochemistry (IHC) was utilized to examine cellular organization within the samples and conditioned media supernatants analyzed for net gelatinase activity. BAM contraction was significantly increased with co-culture. The same side co-culture configuration lead to a greater reduction in surface area than opposite side co-culture. IHC revealed enhanced SMC infiltration into BAM when co-culture was utilized. A significant increase in net gelatinase activity was also observed with the co-culture configuration. Enhanced infiltration and contractile ability of bladder SMCs with UEC co-culture may, in part, be due to an increase in gelatinase activity. The influence of bladder UECs on SMC behaviour in vitro indicates that BAM may contain some key inductive factors that serve to promote important bladder cell-cell and cell-matrix interactions.
Department of Chemical Engineering and Applied Chemistry and the Institute for Biomaterials and Biomedical Engineering, University of Toronto, 200 College Street, Toronto, Ont., Canada M5S 3E5.
Allison L
AL
Brown
Tamara T
TT
Brook-Allred
Jennifer E
JE
Waddell
Jacinta
J
White
Jerome A
JA
Werkmeister
John A M
JA
Ramshaw
Dàrius J
DJ
Bagli
Kimberly A
KA
Woodhouse
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D003902:Detergents / Q000494:pharmacology; D004798:Enzymes / Q000494:pharmacology; D005109:Extracellular Matrix; D005260:Female; D005355:Fibrosis; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D008855:Microscopy, Electron, Scanning; D009130:Muscle, Smooth / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D034421:Sus scrofa; D023822:Tissue Engineering / Q000379:methods*; D001743:Urinary Bladder / Q000033:anatomy & histology* / Q000187:drug effects; D019459:Urothelium / Q000166:cytology* / Q000201:enzymology
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D003902:Detergents; D004798:Enzymes; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9