Cd133 prom1

The presence of CSC surface marker antigens was detected using antibodies CD90/THY1 (Stem Cell Technologies, Vancouver, BC, Canada), CD133/PROM1 (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany), and CD44 (Abcam, Cambridge, UK). Flow cytometric analysis was performed immediately on a FACSCalibur flow cytometer (Becton Dickinson, NJ, USA). A total of 10,000 events were analyzed per sample. Data were presented as a mean for at least three independent experiments.

The presence of surface marker antigens was detected using antibodies CD90/THY1 (Stem Cell Technologies, Vancouver, BC, Canada), CD133/PROM1, CD45 (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany), CD44 (Abcam, Cambridge, UK), and STRO-1 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). After detachment, at least two million cells per mL were incubated with specific first antibodies for 60 minutes on ice in the dark. After two washings with PBS containing 0.5% bovine serum albumin (BSA), when necessary, the cells were incubated with fluorescence-conjugated secondary antibody for 60 minutes on ice in the dark. Flow cytometric analysis was performed immediately in a FACSCalibur flow cytometer (Becton Dickinson, NJ, USA). Ten thousands events were analysed per sample.
The presence and localization of the proteins CD90, CD44, Vimentin/Vim (Abcam), and alpha smooth muscle actin/αSMA (Dako, Glostrup, Denmark) were visualized with immunofluorescence using fluorescence microscope Leica DM2000 (Leica Camera AG, Solms, Germany). The nucleus of the cells was stained with Hoechst 33342 dye (Sigma-Aldrich).