Background: New artificial transplants for reconstruction of longer defects in the urinary tract are still needed. The amnion membrane provides a natural, water-impermeable and mechanically stable basal membrane with several incorporated cell growth promoting factors. Therefore, amnion could represent promising components for scaffolds in artificial urinary transplants. The aim of our study was to define suitable cell and tissue culture conditions for application of amnion membrane (AM) in biological constructs for reconstruction of the urinary tract.
Methods: Several cell culture conditions were examined for the proper isolation of fibroblasts and urothelial cells from porcine urinary bladder. After successful cell culture we seeded both cell types on native and de-epithelized AM. After 15 and 30 days we analyzed cell formation by HE and immunofluorescence staining.
Results: Cultivation of 1cm² tissue specimens with supplemented DMEM-10%FBS led to a positive selection of urothelial cells whereas RPMI with 10%FBS was the medium of choice for successful isolation of fibroblasts. At 15-days, decellularized AM showed insufficient cellular attachment on the major parts of the surface. In native amniotic membranes however, urothelial cells and fibroblasts developed a well differentiated multi- layer cell formation after seeding.
Conclusion: AM is a suitable membrane, giving urothelial cells and fibroblasts ideal conditions. In our experiments, native AM showed better characteristics for cell growing, compared to decellularized AM.
Background: New artificial transplants for reconstruction of longer defects in the urinary tract are still needed. The amnion membrane provides a natural, waterimpermeable and mechanically stable basal membrane with several incorporated cell growth promoting factors. Therefore, amnion could represent promising components for scaffolds in artificial urinary transplants. The aim of our study was to define suitable cell and tissue culture conditions for application of amnion membrane (AM) in biological constructs for reconstruction of the urinary tract. Methods: Several cell culture conditions were examined for the proper isolation of fibroblasts and urothelial cells from porcine urinary bladder. After successful cell culture we seeded both cell types on native and de-epithelized AM. After 15 and 30 days we analyzed cell formation by HE and immunofluorescence staining. Results: Cultivation of 1cm² tissue specimens with supplemented DMEM-10%FBS led to a positive selection of urothelial cells whereas RPMI with 10%FBS was the medium of choice for successful isolation of fibroblasts. At 15-days, decellularized AM showed insufficient cellular attachment on the major parts of the surface. In native amniotic membranes however, urothelial cells and fibroblasts developed a well differentiated multi- layer cell formation after seeding. Conclusion: AM is a suitable membrane, giving urothelial cells and fibroblasts ideal conditions. In our experiments, native AM showed better characteristics for cell growing, compared to decellularized AM.