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Novel method for the isolation and characterisation of the putative prostatic stem cell
Rupesh I Bhatt1, Michael D Brown, Claire A Hart
1Genito-Urinary Cancer Research Group, Cancer Research UK Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
Summary
Researchers identified a side population (SP) of prostate stem cells using Hoechst 33342 dye. This method aids in isolating prostate stem cells, crucial for understanding benign prostate hyperplasia and prostate cancer.
Area of Science:
- Urology
- Stem Cell Biology
- Cell Biology
Background:
- Prostate stem cells are vital for prostate development and function.
- Dysregulation of prostate stem cells is linked to benign prostate hyperplasia (BPH) and prostate cancer (CaP).
- A definitive marker for prostate stem cells has been lacking, hindering research.
Purpose of the Study:
- To adapt a differential dye uptake protocol for isolating putative stem cells from prostate tissue.
- To identify a reliable method for prostate stem cell isolation.
Main Methods:
- Prostate epithelial cells from BPH patients were stained with Hoechst 33342.
- Flow cytometry was used to analyze the Hoechst 33342 Red/Blue profile.
- Cell cycle status of the isolated population was determined using Hoechst 33342 and Pyronin Y staining.
Main Results:
- A verapamil-sensitive side population (SP) was successfully isolated from primary prostate tissue.
- This SP constituted approximately 1.38% of prostate epithelial cells.
- Cell cycle analysis showed the majority of SP cells were in the G0 or G1 phases.
Conclusions:
- The Hoechst 33342 dye efflux protocol is effective for isolating a side population from primary prostate tissue.
- This method provides a means to isolate potential prostate stem cells.