Journal: Nature Communications

Article Title: Human muscle-derived CLEC14A-positive cells regenerate muscle independent of PAX7

doi: 10.1038/s41467-019-13650-z

Figure Lengend Snippet: a PAX7null, PAX7neg, and PAX7pos cells, used in transplantation experiments shown in b , indicate the same differentiation potential (skeletal myosin staining, Fi: Fusion index, Scale bars: 50 µm). b TA muscle sections from NOG mice demonstrate that 3 weeks after transplantation the PAX7null, PAX7neg, and PAX7pos human cell types contributed to muscle regeneration. Human anti-lamin A/C antibody (yellow) detects only human nuclei and anti-human spectrin antibody is specific for fibers of human origin (red). Nuclei in blue (Hoechst). Scale bar: 50 µm. c Quantification of experiments shown in b . Each dot represents one mouse grafted with the cells indicated below ( n = 4 for PAX7null and n = 7 for PAX7neg). Shown are the values for the section with the highest number of human fibers for each mouse and the means. Only human muscle fibers with a diameter > 10 µm were counted. Mice with > 20 human fibers/section were additionally analyzed for PAX7 expression. The presence of PAX7-positive satellite cells is indicated by an open circle. The statistical differences between the groups were analyzed by two-tailed t tests, P > 0.05 = NS (not significant). d Repopulation of satellite cell niche after transplantation: In PAX7null transplants, the satellite cell niche is populated by CLEC14A-positive cells (red, upper row). After transplantation of PAX7neg- and PAX7pos cells, the satellite cell niche is populated by PAX7-positive cells (red, middle, and bottom row). Yellow, human lamin A/C; green, laminin; blue Hoechst. Scale bars: 10 µm (upper row) and 5 µm.

Article Snippet: Cells were collected in Skeletal Muscle Cell Growth Medium (SMCGM, Provitro) and centrifuged for 5 min at 200× g . The resulting cell pellet was resuspended in ice-cold sterile-filtered 1% BSA in PBS (staining buffer) and centrifuged again at 200× g for 5 min at 4 °C.

Techniques: Transplantation Assay, Staining, Expressing, Two Tailed Test