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The long‐term culture of porcine urothelial cells and induction of urothelial stratification
Author(s) -
Ludwikowski,
Bo Zhang,
Frey Frey
Publication year - 1999
Publication title -
bju international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.773
H-Index - 148
eISSN - 1464-410X
pISSN - 1464-4096
DOI - 10.1046/j.1464-410x.1999.00221.x
Subject(s) - urothelial cell , term (time) , urothelial cancer , stratification (seeds) , cancer research , biology , medicine , urothelium , urinary bladder , physics , cancer , bladder cancer , seed dormancy , germination , botany , quantum mechanics , dormancy
Objective To assess porcine urothelial cell cultures and the in vitro induction of urothelial stratification in long‐term cultures, to study their morphological, functional and genetic behaviour, and thus provide potential autologous urothelium for tissue‐engineered substitutes for demucosalized gastric or colonic tissue. Materials and methods Primary cultures of porcine urothelium were established and the cells passaged thereafter. Cell specificity was confirmed by cytokeratin analysis, cell membrane stability assessed using lactate dehydrogenase leakage, cell de‐differentiation by γ‐glutamyl transferase activity and genomic stability by karyotype investigations. Histology and scanning electron microscopy were performed to study the cultured cells and the stratified constructs. Furthermore, collagen matrices were tested as cell scaffolds. Results The cells were cultured for 180 days; 10 subcultures were established during this period. Stratification was induced in a culture flask and on a collagen matrix. Cytokeratins 7, 8, 17 and 18 were expressed in all cultures, and cell membranes were stable, with no evident de‐differentiation. The cultures were stable in their genotype and no chromosomal aberrations were found. The histology and immunohistochemistry of the stratified porcine constructs, and cell membrane stability and cell de‐differentiation, were compared with those in the human system. Conclusion Pig and human urothelial cells can be cultured over a long period with no signs of senescence. Urothelial stratification can be induced in vitro . The collagen matrix seems to be an excellent scaffold, allowing cell adherence and growth.