Figure 4.

Expression of myogenic-specific genes in differentiated menstrual blood–derived cells. Menstrual blood–derived cells were cultured in DMEM supplemented with 20% FBS, 2% HS, or serum-free ITS medium. (A) RT-PCR analysis with PCR primers that allows amplification of the human MyoD, Myf5, desmin, myogenin, MyHC-IIx/d, and dystrophin cDNA (from top to bottom). RNAs were isolated from menstrual blood–derived cells in DMEM supplemented with 20% FBS at the indicated day after treatment with 5 μM 5-azacytidine for 24 h. RNAs from human muscle and H₂O served as positive (P) and negative (N) controls. Only the 18S PCR primer reacted with the human and murine cDNA. (B) Immunocytochemical analysis using an antibody to desmin (a–f) was performed on the menstrual blood–derived cells at 2 wk after exposure to 5 μM of 5-azacytidine for 24 h. The desmin-positive cells are shown at higher magnification (d–f). Merge of a and b is shown in c, and merge of d and e is shown in f. The images were obtained with a laser scanning confocal microscope. Scale bars, 200 μm (a–c) and 75 μm (d—f). (C and D) RT-PCR analysis of menstrual blood–derived cells on DMEM supplemented with 2% HS (C) or serum-free ITS medium (D) at the indicated day after exposure to 5 μM 5-azacytidine for 24 h. (E–G) Western blot analysis was performed on the cells cultured in myogenic medium indicated for 21 d. The blot was stained with desmin (E), myogenin (F), and dystrophin (G) antibodies followed by an HRP-conjugated secondary antibody.