Document Detail

Transplantable urothelial cell sheets harvested noninvasively from temperature-responsive culture surfaces by reducing temperature.
MedLine Citation:
PMID:  14633384     Owner:  NLM     Status:  MEDLINE    
Augmentation cystoplasty using gastrointestinal flaps may induce severe complications such as lithiasis, urinary tract infection, and electrolyte imbalance. The use of viable, contiguous urothelial cell sheets cultured in vitro should enable us to avoid these complications. Transplantable urothelial cell sheets were obtained by utilizing a temperature-responsive cell culture method, and then examined by immunostaining and electron microscopy. Canine urothelium was produced on the surfaces of temperature-responsive culture dishes covalently bonded with the thermally sensitive polymer, poly(N-isopropylacrylamide). Stratified urothelial cell sheets were cultured and then harvested intact without enzymatic treatment from these dishes by reducing the temperature. Histological structure and cell-to-cell junctions were compared between these urothelial cell sheets and those harvested with dispase. All urothelial cell sheets were harvested from the bonded surfaces by reducing the culture temperature without the need for dispase. Electron microscopy revealed well-developed microridge, microvilli, and cell junction complexes. Conversely, these same cell features were destroyed by dispase treatment. Immunoblotting revealed that dispase fragmented occludin, whereas it remained unchanged in the intact urothelial cell sheets. Novel urothelial cell sheets obtained by culture on temperature-responsive culture surfaces were successfully harvested much less destructively than with dispase. This technology should prove useful in urinary tract tissue engineering in the near future.
Yoshiyuki Shiroyanagi; Masayuki Yamato; Yuichiro Yamazaki; Hiroshi Toma; Teruo Okano
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Tissue engineering     Volume:  9     ISSN:  1076-3279     ISO Abbreviation:  Tissue Eng.     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2003-11-24     Completed Date:  2004-06-08     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9505538     Medline TA:  Tissue Eng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1005-12     Citation Subset:  IM    
Institute of Advanced Biomedical Engineering and Science and Department of Urology, Tokyo Women's Medical University, Tokyo, Japan.
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MeSH Terms
Cell Culture Techniques
Cold Temperature*
Gastrointestinal Diseases / surgery,  therapy
Microscopy, Electron
Staining and Labeling
Urothelium / transplantation*,  ultrastructure

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