Premium
Novel method for the isolation and characterisation of the putative prostatic stem cell
Author(s) -
Bhatt Rupesh I.,
Brown Michael D.,
Hart Claire A.,
Gilmore Paul,
Ramani Vijay A.C.,
George Nicholas J.,
Clarke Noel W.
Publication year - 2003
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.10058
Subject(s) - side population , prostate , stem cell , prostate cancer , population , hyperplasia , biology , pathology , cancer research , medicine , cancer stem cell , microbiology and biotechnology , cancer , environmental health
Background Prostate stem cells, responsible for the development, maturation, and function of the prostate, have been implicated in the aetiology of both benign prostate hyperplasia (BPH) and prostate cancer (CaP). However, research has been hampered by the lack of a definitive stem cell marker. We have adapted the protocol for differential Hoechst 33342 uptake by hemopoietic stem cells to enable isolation of putative stem cells from the prostate. Methods Prostate epithelial cells isolated from prostate tissue obtained from patients with BPH after transurethral resection of the prostate were stained with Hoechst 33342. The Hoechst 33342 Red/Blue flow cytometry profile was then determined. Hoechst 33342 and Pyronin Y staining was used to determined the cell cycle status. Results A verapamil‐sensitive side population (SP) can be isolated from primary prostate tissue accounting for 1.38% ± 0.07% of prostate epithelial cells. Cell cycle analysis of this SP population revealed that the majority of SP cells are in either G 0 (12.38 ± 0.31%) or G 1 (63.19 ± 2.13%). Conclusions The Hoechst 33342 dye efflux protocol can be adapted for the isolation of a SP from primary prostate tissue. Cytometry Part A 54A:89–99, 2003. © 2003 Wiley‐Liss, Inc.