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A Single Cell Dissociation Approach for Molecular Analysis of Urinary Bladder in the Mouse Following Spinal Cord Injury
Published on: June 17, 2020
Do current bladder smooth muscle cell isolation procedures result in a homogeneous cell population? Implications for
Arun K Sharma1, Jena L Donovan, Jennifer A Hagerty
1Department of Urology, The Feinberg School of Medicine at Northwestern University, IBNAM 11-113, Chicago, IL 60611, USA. arun-sharma@northwestern.edu
World Journal of Urology
|February 24, 2009
Summary
Conventional bladder smooth muscle cell (SMC) culturing methods yield mixed cell populations, not uniform SMCs. This heterogeneity impacts bladder tissue engineering applications, necessitating standardized characterization criteria.
Area of Science:
- Urology
- Cell Biology
- Tissue Engineering
Background:
- Smooth muscle cells (SMCs) are crucial for bladder function.
- Conventional methods for isolating and culturing bladder SMCs are widely used.
- The homogeneity of cultured bladder cells is often assumed for tissue engineering.
Purpose of the Study:
- To investigate the cellular composition of bladder-derived cells obtained through conventional harvesting and culturing techniques.
- To determine if current methods yield a uniform population of bladder SMCs suitable for tissue engineering.
Main Methods:
- Patient-derived bladder muscle was isolated, minced, and enzymatically digested.
- Cells were cultured and analyzed using flow cytometry and immunofluorescence.
- Antibodies targeting SMC and non-SMC markers (alpha-SMA, vWF, pan-cytokeratin, CD31, CD90) were employed.
Main Results:
- Flow cytometry and immunofluorescence revealed mixed cell populations expressing non-SMC markers.
- Oligonucleotide array analysis confirmed the presence of endothelial and fibroblast markers alongside SMC genes.
- Significant expression of non-SMC markers was observed (P < 0.005).
Conclusions:
- Conventional methods for acquiring and culturing bladder SMCs result in heterogeneous cell populations.
- The presence of non-SMC markers challenges the assumption of homogeneity.
- Standardized criteria are needed to accurately characterize bladder SMCs for tissue engineering.
