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. Author manuscript; available in PMC: 2015 Mar 16.
Published in final edited form as: J Orthop Res. 2013 Apr 1;31(7):1089–1095. doi: 10.1002/jor.22335

Figure 1.

Figure 1

Characterization of clonal myogenic endothelial cells (cMECs) in culture. (A) RT-PCR analysis was performed on all six FACS-sorted MEC clones and compared with HUVECs, cultured unsorted hPSMCs (Unsorted), and fresh skeletal muscle cell lysate (Fresh total cells). All MEC clones consistently expressed myogenic (desmin, CD56, Pax7, m-cadherin, Pax3 Myf5), endothelial (CD34, VE-cadherin, von Willebrand Factor (vWF)), smooth muscle/vascular mural (α-smooth muscle actin, PDGFR-β, NG2, CD146), and mesenchymal stem/stromal (CD90 and CD105) cell markers. (B) Analysis of cMEC clonogenic proliferation capacity by sub-cloning single cells from GFP-transduced MEC clones. A total of 80 sub-cloned single GFP-positive cMEC cells (31% of 258 seeded wells) were tracked. Among them, 1% died, 12% did not divide, 14% divided into two to four cells, and 73% were able to form colonies (>8 cells). (C) Colony growth rate (n = 4) after 17 days in culture showed that the population doubling time was 28.1 ± 5.5 h, and the cell division time was 16.8 ± 2.1 h. (D) After 10 passages in culture, the majority of cMEC metaphases analyzed possess an euploid number (46) of chromosomes.