Figure 1 | Scientific Reports

Figure 1

From: Direct reprogramming of fibroblasts into skeletal muscle progenitor cells by transcription factors enriched in undifferentiated subpopulation of satellite cells

Figure 1

Separation of functionally distinct subpopulation of primary cultured satellite cells by Calcein-AM. (a) Representative fluorescence images of calcein-AM-treated cSCs 7 days after isolation. Upper left images show strong fluorescence in differentiated myotubes. Arrow and arrowhead show differentiated and undifferentiated cells, respectively. Upper right and lower images show heterogeneous fluorescence in round, undifferentiated cells in low-power and high-power fields, respectively. n > 3. Scale bars: 500 µm upper, 100 µm lower. (b) Representative FACS analysis of calcein-AM-treated C2C12 myoblasts (upper) and cSCs 7 days after isolation. The lower, middle, and upper 10–15% of the population were sorted as Calceinlow, Calceinmiddle, and Calceinhigh cSCs, respectively. n > 3. (c) Expression of Pax7, Myf5, MyoD, Myogenin, and Desmin in Calceinlow, Calceinmiddle, and Calceinhigh cSCs. n = 6–8. (d) Representative bright-field images of Calceinlow, Calceinmiddle, and Calceinhigh cSCs 0 and 3 days after FACS sorting. n = 4. Scale bar: 200 µm. (e) Quantitative analysis of numbers of Calceinlow, Calceinmiddle, and Calceinhigh cSCs. n = 4. (f) Representative images of BrdU-positive Calceinlow, Calceinmiddle, and Calceinhigh cSCs. n = 5. Scale bar: 200 µm. (g) Quantitative analysis of BrdU-positive cells. n = 5. (h) Representative images of Dystrophin-positive myofibers in mdx mice transplanted with Calceinlow, Calceinmiddle, and Calceinhigh cSCs, respectively. Scale bar: 100 µm. (i) Quantitative analysis of numbers of Dystrophin-positive fibers in transplanted tibialis anterior/extensor digitorum longus muscles. n = 3. Error bars indicate SEM. *P < 0.05, **P < 0.01, ***P < 0.001 by ANOVA with Tukey–Kramer test.

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