Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro

B. G. Cilento, M. R. Freeman, F. X. Schneck, A. B. Retik, A. Atala

Research output: Contribution to journalArticle

179 Citations (Scopus)

Abstract

A simple method for the harvest of bladder cell types from surgical specimens was used to generate strains of normal human urothelial cells that could be reproducibly cultivated, passaged and extensively expanded in serum- free medium. Immunostaining of the bladder epithelial cells with broadly reacting anti-cytokeratin antibodies and with an anti-cytokeratin antibody specific to cytokeratin 7, a transitional cell marker, indicated that they expressed a stable epithelial phenotype with serial passage. Low levels of immunostaining for E-cadherin and low levels of E-cadherin messenger ribonucleic acid, as determined by Northern blot analysis, and strongly positive immunostaining with an anti-vimentin antibody indicated collectively that the uroepithelial cells express a nonbarrier-forming phenotype under these culture conditions. However, when the urothelial cells were implanted subcutaneously into athymic mice on biodegradable synthetic polymers, they formed multilayered structures, suggesting that they retain the capability to differentiate in a living host. The urothelial cells proliferated in an epidermal growth factor independent manner and expressed high levels of transforming growth factor-α and amphiregulin messenger ribonucleic acids, suggesting the possibility of autocrine regulation of growth by epidermal growth factor-like factors. Cytogenetic analysis indicated that urothelial cells cultured for 6 passages possessed a normal chromosomal complement. These results demonstrate that primary cultures of autologous human bladder epithelial cells can be extensively expanded in vitro and, consequently, might be used in cell transplantation strategies for genitourinary reconstruction.

Original languageEnglish
Pages (from-to)665-670
Number of pages6
JournalJournal of Urology
Volume152
Issue number2 II
StatePublished - Jan 1 1994
Externally publishedYes

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Urothelium
Cytogenetics
Urinary Bladder
Anti-Idiotypic Antibodies
Cadherins
Keratins
Epidermal Growth Factor
Epithelial Cells
RNA
Keratin-7
Serial Passage
Phenotype
Cytogenetic Analysis
Cell Transplantation
Serum-Free Culture Media
Transforming Growth Factors
Vimentin
Nude Mice
Northern Blotting
In Vitro Techniques

Keywords

  • amphiregulin
  • bladder
  • cytogenetics
  • phenotype
  • transforming growth factor alpha

ASJC Scopus subject areas

  • Urology

Cite this

Cilento, B. G., Freeman, M. R., Schneck, F. X., Retik, A. B., & Atala, A. (1994). Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro. Journal of Urology, 152(2 II), 665-670.

Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro. / Cilento, B. G.; Freeman, M. R.; Schneck, F. X.; Retik, A. B.; Atala, A.

In: Journal of Urology, Vol. 152, No. 2 II, 01.01.1994, p. 665-670.

Research output: Contribution to journalArticle

Cilento, BG, Freeman, MR, Schneck, FX, Retik, AB & Atala, A 1994, 'Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro', Journal of Urology, vol. 152, no. 2 II, pp. 665-670.
Cilento BG, Freeman MR, Schneck FX, Retik AB, Atala A. Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro. Journal of Urology. 1994 Jan 1;152(2 II):665-670.
Cilento, B. G. ; Freeman, M. R. ; Schneck, F. X. ; Retik, A. B. ; Atala, A. / Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro. In: Journal of Urology. 1994 ; Vol. 152, No. 2 II. pp. 665-670.
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