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First published online 29 June 2004
doi: 10.1242/jcs.01222

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CD133, a novel marker for human prostatic epithelial stem cells

¹ Prostate Research Group, Surgical Oncology, Medical School, Framlington Place, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH, UK
² YCR Cancer Research Unit (Area 13), Department of Biology, University of York, PO BOX 373, York, YO10 5DD, UK
³ Department of Oncology, Addenbrooke's Hospital, University of Cambridge, Hills Road, Cambridge, CB2 2QQ, UK

View larger version (115K): [in a new window]   Fig. 1. A rare sub-set of basal cells express CD133+. A paraffin section of prostatic acini labelled with the nuclear stain DAPI (blue) and anti-CD133 directly conjugated to PE (red). 200 cross sections of acini were studied by confocal microscopy for the presence and location of CD133+ cells. Scale bar: 40 µm.

View larger version (30K): [in a new window]   Fig. 2. CD133+ cells are restricted to the 2ß1hi population and are largely quiescent. Basal cells were selected for 2ß1hi and 2ß1low expression. (A) 2ß1hi population labelled with antibodies to CD133 (red) and basal cell marker 34ßE12 (green). Co-localization (yellow/orange; yellow arrow). Scale bar: 5 µm. (B,C) Graph of 2ß1hi population (B) and 2ß1low population (C) labelled with anti-CD133 antibody (shaded area) and analysed by flow cytometry. Isotype control is depicted as a solid black line. (D) Dot-plot showing flow cytometric analysis of basal cells double-labelled with anti-CD133 (APC) and Ki-67 (FITC) from a representative experiment (n=3).

View larger version (10K): [in a new window]   Fig. 3. 2ß1hi/CD133 population have a high proliferative potential in vitro. (A) Basal cells with the phenotypes 2ß1hi/CD133+ and 2ß1hi/CD133- were plated onto type I collagen plates and the colony forming efficiency (CFE) determined. Cells were counted before the addition of irradiated feeders and were cultured for up to 28 days. Controls (basal cells) were plated with no pre-selection. Colonies containing 32 or more cells were scored. CFE was calculated as the number of colonies formed per number of selected cells x100. CFEs are expressed as the ratio of the selected population to control (unselected basal cell) population. Results show means ± s.e.m. of four experiments. (B) Long-term proliferative potential of 2ß1hi/CD133+ and 2ß1hi/CD133- cells. 5x103 cells were seeded onto type I collagen plates, in triplicate, and the total cell output was determined at the end of the serial culture when their growth capacity was exhausted. Results show means ± s.e.m. of four experiments. P<0.05.

View larger version (50K): [in a new window]   Fig. 4. Phenotype of 2ß1hi/CD133+ basal cells. 2ß1hi/CD133+ basal cells were isolated, plated on to collagen I-coated culture slides and triple labelled with anti-keratin antibodies and the nuclear stain, DAPI. (A) 34ßE12 (green). (B) K5 (green) and K14 (red). (C) K18 (green) and K19 (red). (D) K14 (red) and K19 (green). (E) K14 (red) and K18 (green). Approximately 100 cells were isolated for each experiment (n=10). Scale bar: 10 µm.

View larger version (127K): [in a new window]   Fig. 5. CD133+ selected cells form fully differentiated acini in immunocompromised mice. Xenografts of prostate acini formed by transplantation of 2ß1hi/CD133+ basal cells stained with (A) Haematoxylin and Eosin, (B) 34ßE12, (C) anti-K18, (D) anti-PAP (E) Anti-androgen receptor. Scale bar: 40 µm.