Characterization of BMSSCs in vivo.

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Characterization of BMSSCs in vivo. Characterization of BMSSCs in vivo. (A) Light microscopic examination of cytospins representing freshly sorted STRO-1BRIGHT/VCAM-1+ marrow cells (1000×) counterstained with heamatoxylin. (B) Transmission electron micrograph depicting the ultrastructure of freshly sorted STRO-1BRIGHT/VCAM-1+ cells (15000×). Immunohistochemical staining of cytospin preparations of the sorted STRO-1BRIGHT/VCAM-1+ cells (1000×) with collagen type I (C) and α-SMA (D). (E) Dual-color flow cytometric analysis of Ki67 (FITC) expression by STRO-1+ (PE) marrow cells isolated by MACS. The majority of the STRO-1BRIGHT marrow cells lack detectable binding of Ki67 relative to that of the isotype-matched control antibody, indicated by the vertical quad-stat marker. (F) Telomerase activity in different sorted cell populations was examined using a modified TRAP assay. TRAP products derived from CHAPS extracts of non-denatured (-) and denatured (+) total BM (lanes 1), STRO-1BRIGHT/VCAM-1+ cells sorted fraction (lanes 2) and CD34+-sorted BM cells (lanes 3). TRAP products were resolved on a 12% polyacrylamide gel, stained with SYBR green fluorescent dye, and visualised using a fluorescence scanning system. (G) A typical light microscopic view of a single purified STRO-1BRIGHT/VCAM-1+ cell, allowed to adhere to fibronectin-coated culture plates (400×). (H) A representative example of a day 14 CFU-F colony stained with toluidine blue is shown (200×). (I) Immunohistochemical staining showing all the cells that comprise the CFU-F colony express collagen type I (200×). Stan Gronthos et al. J Cell Sci 2003;116:1827-1835 © The Company of Biologists Limited 2003