Expansion and long-term culture of differentiated normal rat urothelial cells in vitro

Détails

ID Serval
serval:BIB_5EA02FA16AD5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Expansion and long-term culture of differentiated normal rat urothelial cells in vitro
Périodique
In Vitro Cellular and Developmental Biology Animal
Auteur(s)
Zhang  Y. Y., Ludwikowski  B., Hurst  R., Frey  P.
ISSN
1071-2690
Statut éditorial
Publié
Date de publication
08/2001
Peer-reviewed
Oui
Volume
37
Numéro
7
Pages
419-29
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul-Aug
Résumé
The objective of this study is to establish a reliable cell culture system for the long-term culture of rat urothelial cells (RUC), in which the cells multiply in vitro and form stratified polarized urothelium. Urothelial cells were harvested by the enzymatic digestion of the urothelium exposed by the eversion of resected rat bladders. Primary cultures were initiated in keratinocyte serum-free medium (KSFM) for selective proliferation of urothelial cells. Subsequently, the cells were propagated in a mixture of conditioned medium (CM) derived from Swiss 3T3 cell culture supernatant and KSFM (CM-KSFM). Mean population doubling time was 13.8 +/- 0.9 h. RUC were successfully maintained for 18 passages over a period of 4-5 mo. Detailed investigations of culture conditions showed that CM-KSFM yielded a differentiated multilayer structure. The stratified urothelial sheets measuring 4 x 6 cm2 could be formed and then detached using dispase. Cytokeratin pattern in both the cultured urothelial monolayer and engineered stratified layers was similar to those seen in vivo, as assessed with monoclonal antibody against cytokeratin 17. Ultrastructural morphology showed microvilli, basal cell layer, and desmosomes between adjacent cells in the stratified urothelium.
Mots-clé
3T3 Cells Animals Antibodies, Monoclonal *Cell Differentiation *Cell Division Cells, Cultured Culture Media, Conditioned Culture Media, Serum-Free Immunohistochemistry Keratins/analysis Male Mice Microscopy, Electron Rats Rats, Wistar Time Factors Urothelium/*cytology
Pubmed
Web of science
Création de la notice
28/02/2008 11:25
Dernière modification de la notice
20/08/2019 15:16
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