In Vivo Tracking of Murine Adipose Tissue-Derived Multipotent Adult Stem Cells and Ex Vivo Cross-Validation

Figure 1

mAT-MASC^eGFP characterization. (a) Phase contrast (upper left) and eGFP fluorescence (lower left) images of mAT-MASC^eGFP at the third passage in culture. The right panel represents the overlapping of the previous ones (calibration bar 100 μm). (b) Surface immunophenotype: representative flow cytometry histograms of mAT-MASC. Plots show isotype control IgG-staining profile (green histogram) versus specific antibody staining (red histogram). (c–f) Pluripotent state-specific transcription factor expression: Oct-4 (green fluorescence; (c)), Nanog (red fluorescence; (d)), and Sox-2 expression (yellow fluorescence; (e)) in the nuclei of mAT-MASC. Nuclei are depicted by the blue fluorescence of DAPI staining ((c–e), calibration bars 20 μm). (f) Quantification of pluripotent state-specific transcription factor expression. Data are presented as mean ± standard deviation. (g–k) Multipotency of mAT-MASC^eGFP. mAT-MASC cultured in osteogenic medium display positivity for von Kossa (brown deposits; (g), calibration bar 100 μm), while cultured in adipogenic medium acquired positivity for Oil-Red-O (red deposits; (h), calibration bar 100 μm). A fraction of mAT-MASC^eGFP in myogenic medium expressed the myocyte-specific filament SMA (green fluorescence, (i), calibration bar 20 calibration bar 100 μm) and displayed spontaneous calcium transients (j-k). Specifically, cells loaded with Fluo-4 emitted green fluorescence in response to intracytoplasm calcium release (green fluorescence; (j)). In the graph (k), each colored line represents the changes of fluorescence intensity with time within the ROI identified in Figure (k) by the same color.