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Isolation and characterisation of prostate stem-like cells

Yamamoto, H.; (2012) Isolation and characterisation of prostate stem-like cells. Doctoral thesis , UCL (University College London).

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Emerging evidence suggests that abnormal stem cells play a critical role in cancer development and progression. The aim of the thesis was to identify and characterise benign and malignant stem-like cells from the prostate gland. The central hypothesis tested in this study was that prostate stem-like cells (PSLCs) from benign and malignant prostate glands are identified by cell surface marker expression. Culture systems were compared to establish the conditions for mouse PSLC (mPSLC) culture. A rare stem cell-like subpopulation (0.2-0.8% of total cells) capable of clonal proliferation, self-renewal, and gland-like structure formation were identified within monolayer and spheroid-colony forming cultures, but not in suspension culture. Mouse prostate cells sorted according to expression of one or more candidate cell surface markers showed CD49f+Sca-1+ cells to contain the highest proportion of mPSLCs. mPSLC properties were also investigated in a model of prostate cancer. Cancer-derived cells produced larger spheroid colonies and contained higher proportions of CD49f+Sca-1+ cells than benign controls, suggesting dysregulated PSLC function. Stem-like cells were also detected in the human prostate (hPSLCs) representing 0.5% of total cells. CD49f+ selection resulted in significant enrichment and high recovery of hPSLCs, whereas CD44+ or CD133+ selection led to hPSLC depletion compared to unsorted cells. By immunohistochemistry, CD49f expression was detected in the basal layer, consistent with the predicted niche of stem cells. Using an optimised protocol of cell isolation, hPSLCs were isolated from needle biopsy tissue containing advanced prostate cancer cells. Cancer-derived hPSLCs were best isolated by CD49f+ selection rather than by CD44+ or CD133+ selection, similar to benign PSLCs. The study identified an efficient marker for stem-like cells of the human and mouse prostate. Future studies will identify differences between stem-like cells from cancer and benign tissues as well as novel therapeutic approaches for PSLC targeting in cancer.

Type: Thesis (Doctoral)
Title: Isolation and characterisation of prostate stem-like cells
Language: English
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Surgery and Interventional Sci > Department of Targeted Intervention
URI: https://discovery.ucl.ac.uk/id/eprint/1344100
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