Generation of a Functional, Differentiated Porcine Urothelial Tissue In Vitro

The Distribution and Function of Chondroitin Sulfate and Other Sulfated Glycosaminoglycans in the Human Bladder and Their Contribution to the Protective Bladder Barrier

“…26 Although our results revealed distinct differences between the locations of different GAGs between the urothelial luminal surface and the underlying urothelium layers, slight contamination . Note percent decrease in TEER compared to baseline using Hep3 enzymes to digest HS and chondroitin ABCase (CSabc-ase) enzymes to digest CS.…”

“…1,21 The results of this study demonstrate that high TEER can be achieved by the applied culturing methods, although there were some differences in applied cell culturing protocols compared to those of others. 26 Urothelial TEER values in our study were comparable to those of bladder tissue from other mammals 29,30 and could be considered a tight epithelium according to the classification of Fromter and Diamond. 21 Protamine, which was chosen as a positive control for TEER experiments, causes a barrier defect by eliminating all GAGs and creating mild urothelial cell damage.…”

Tissue Engineered Tubular Construct for Urinary Diversion in a Preclinical Porcine Model

“…For enzymatic treatment the biopsy tissue was placed in stripping medium composed of HBSS without Ca 2ϩ and Mg 2ϩ , 10 mM HEPES, 0.1% aprotinin, 1% penicillin/streptomycin and 2.4 U/ml dispase II overnight at 4C. 16 The urothelium was removed using forceps and digested in collagenase IV solution composed of HBSS with Ca 2ϩ and Mg 2ϩ , 10 mM HEPES and 100 U/ml collagenase IV for 20 minutes at 37C. UCs were suspended and collected in keratinocyte serum-free medium (Invitrogen™) with 0.5 ng/ml epidermal growth factor, 5 ng/ml bovine pituitary extract, 30 ng/ml cholera toxin, 100 U penicillin per ml and 100 g streptomycin per ml PBS, centrifuged, resuspended and cultured on a mouse STO fibroblast feeder layer in T75 Primaria™ flasks.…”

Microstructure and cytocompatibility of collagen matrices for urological tissue engineering

“…To accomplish such stratification, the use of a specific induction medium containing high calcium concentrations [13,14] or keratinocyte growth factor (also known as fibroblast growth factor 7) [15] is needed. However, these media are not dedicated for smooth muscle cell expansion.…”