Document Detail

Increased Urothelial Cell Detection in the Primary Bladder Smooth Muscle Cell Cultures with Dual MACS/qRT-PCR Approach.
MedLine Citation:
PMID:  20930617     Owner:  NLM     Status:  In-Data-Review    
Bladder tissue has been regenerated in humans with neurogenic bladder using an implant produced from autologous urothelial (UC) and smooth muscle cells (SMC) expanded from bladder biopsies seeded onto a biodegradable synthetic scaffold. As the majority of bladder cancers are urothelial carcinomas (aka, transitional cell carcinoma), this 2-cell type autologous sourcing strategy presents significant challenges to product development. Entire bladders have been regenerated in cystectomized animals using a single-cell-type sourcing strategy: implants were seeded with bladder-derived SMC-only. Applying the bladder SMC-only sourcing strategy to produce clinical implants for bladder replacement or urinary diversion in bladder cancer patients requires methods for screening SMC cultures for the presence of potentially cancerous UC cells to provide evidence of SMC culture purity before seeding the scaffold. In this report, we show a 10-fold to 100-fold improvement in the sensitivity of qualitative and quantitative reverse-transcription PCR (qRT-PCR)-based assays for detecting UC positive for Cytokeratin 5 (CK5) in mixed SMC/UC cultures when the cell population was first subjected to magnetic activated cell sorting to enrich for cells expressing the epithelial cell adhesion molecule (known as EPCAM or CD326), a marker known to be present in normal UC and upregulated in the cancerous UC.
Chistopher W Genheimer; Kelly I Guthrie; Jacob E Shokes; Andrew T Bruce; Sarah F Quinlan; Namrata Sangha; Roger M Ilagan; Joydeep Basu; Teresa Burnette; John W Ludlow
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Applied immunohistochemistry & molecular morphology : AIMM / official publication of the Society for Applied Immunohistochemistry     Volume:  19     ISSN:  1533-4058     ISO Abbreviation:  Appl. Immunohistochem. Mol. Morphol.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-02-10     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100888796     Medline TA:  Appl Immunohistochem Mol Morphol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  184-9     Citation Subset:  IM    
Bioprocess Research and Assay Development, Tengion Inc, Winston-Salem, NC.
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