Premium
In vitro assessment of a collagen sponge for engineering urothelial grafts
Author(s) -
Sabbagh,
Masters,
Duffy,
Herbage,
Brown
Publication year - 1998
Publication title -
british journal of urology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.773
H-Index - 148
eISSN - 1464-410X
pISSN - 0007-1331
DOI - 10.1046/j.1464-410x.1998.00828.x
Subject(s) - sponge , urothelial cell , immunostaining , cell culture , cell growth , in vitro , tissue engineering , cell , chemistry , calcium , pathology , biology , anatomy , urothelium , biomedical engineering , immunohistochemistry , medicine , urinary system , biochemistry , botany , genetics , organic chemistry
Objective To investigate the deposition of urinary crystals and the growth characteristics of urothelial cells on a collagen sponge, as a preliminary step in engineering urothelial autologous grafts. Materials and methods Collagen sponges were exposed to a continuous flow of urine at pH 5.3 and 6.3 for 1 week. The sponges were examined microscopically for crystal deposition and analysed for their calcium content. Two cell lines, RT112, derived from a well‐differentiated transitional cell carcinoma, and UROtsa, an immortalized urothelial cell line, were seeded on the collagen sponges. Cells were cultured for 6, 12 and 21 days. The pattern of growth was analysed by histology and immunostaining with a pan‐cytokeratin antibody. Growth was assayed to quantify cell proliferation on the sponges. Results No crystals were evident on any of the collagen sponges. Calcium deposition was negligible at pH 5.3. Although calcium levels were measurable at pH 6.3, the levels were very low. Both cell lines attached and grew in a stratified manner on the collagen sponge, RT112 forming a layer 6–8 cells thick, and UROtsa a layer 4–6 cells thick; cell proliferation was maximal at 5–10 days. The sponge remained easy to handle after 3 weeks in culture. Conclusion These findings show that collagen sponges support the growth and stratification of urothelial cells, and indicate that the collagen sponge is a suitable substrate for developing urothelial autologous grafts.