Fig. 1. Multipotentiality of MMCs derived from muscle satellite cells. (A-D) Expression of myogenic lineage markers in a clone (GB1T) of primary cultured MMCs was determined by immunofluorescence analysis with antibodies to MyoD (A), to Pax7 (B), to desmin (C) or to nestin (D), as well as Cy3-conjugated secondary antibodies. (E) Undifferentiated MMCs resembled fibroblasts when cultured in pmGM. (F) MMCs differentiated into myotubes expressing sarcomeric myosin heavy chain (immunostained with a horseradish peroxidase reaction product) after culture for 4 days in pmDM. (G) MMCs differentiated into immature osteoblasts expressing ALP (activity detected by staining with Fast Blue RR) when cultured in pmDM supplemented with BMP2 (250-500 ng ml–1) for 4 days. (H) MMCs differentiated into adipocytes containing many lipid droplets in their cytoplasm (as revealed by staining with oil red O; nuclei were detected by staining with DAPI) when cultured for 6 days in DMEM supplemented with 10% FBS and 100 µM 
-linolenic acid. Images in A-D,H were obtained by epifluorescence microscopy, that in E was obtained by phase-contrast microscopy, and those in F,G were obtained by bright-field microscopy. Scale bars: 50 µm.
