Novel method for the isolation and characterisation of the putative prostatic stem cell

Abstract

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 G0 (12.38 ± 0.31%) or G1 (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.

Normal ovarian surface epithelial label-retaining cells exhibit stem/progenitor cell characteristics

“…The side population phenomena of Hoescht 33342 dye efflux has been used to identify a variety of somatic as well as cancer stem cells in various tissues (20)(21)(22)(23)(24)(25)(26)(27)(28). The ability to efflux a variety of chemicals is postulated to be a defense mechanism, which leads to the longevity required of somatic stem cells and the chemoresistance characteristic of cancer stem cells (29).…”

“…Additionally, somatic and cancer stem cells from various tissues have been identified by their ability to efflux Hoechst 33342 dye through ATP-binding-cassette transporters, such as Abcg2/Bcrp1 (11,(20)(21)(22)(23)(24)(25)(26)(27), including our recent identification of these ''side population'' (SP) cells as potential tumor-initiating cells in ovarian cancer (28). It has been postulated that this chemical-effluxing capability contributes to the cytopreservation necessary for the longevity attributed to stem/progenitor cells (29).…”

Epithelial Stem Cells of the Prostate and Their Role in Cancer Progression

“…ABCG2 was shown to be efficient at Hoechst dye efflux, creating a side population (SP) phenotype that has been used as an alternate approach to identify multipotent stem cells in bone marrow, skeletal muscle, and neural tissue (Bunting 2002) and more recently in the prostate (Bhatt et al 2003). The SP cells of the prostate have slightly higher sphere-forming activity in vitro than the non-SP cells, although only by fourfold (Brown et al 2007).…”

Highly purified CD44+ prostate cancer cells from xenograft human tumors are enriched in tumorigenic and metastatic progenitor cells

“…That the adult human prostate contains stem cells (SCs) is also supported by the observation that a small number of cells within the prostate possess tremendous proliferative capacity and can form glandular-like structures in reconstituted systems (Hudson et al, 2000). Several populations of putative human prostate stem/progenitor cells have been reported, which include CK5 and CK18 double-positive (CK5 þ /CK18 þ ) intermediate cells (van Leenders et al, 2000;Wang et al, 2001;Tran et al, 2002;Garraway et al, 2003;Bhatia et al, 2005), the side population (SP) cells (Bhatt et al, 2003), and small populations of cells that preferentially express cell surface molecules CD44 (Liu et al, 1997), a2b1 (Collins et al, 2001), or CD133 (Richardson et al, 2004).…”