Figure 5

Porcine satellite cells lost their stemness after long-term expansion in vitro. (a) Representative phase contrast images of pig satellite cells cultured in F10 medium for different passages. Scale bars, 200 μm. (b) Growth curves of pig satellite cells isolated from 1-week-old pig. Cells were serially expanded and counted at each passage. (c) FACS analysis of CD56 and CD29 expression from different passages of pig satellite cells cultured in F-10 medium. (d) Quantification of the number of Pax7⁺ cells from different passages. Error bars represented S.E.M. and were based on three independent experiments. (e) qRT-PCR analysis of Pax7 mRNA levels from cells being expanded for different passages. Error bars represented S.E.M. and were based on three independent experiments. (f) Representative immunofluorescent staining images of MyHC staining of myotubes differentiated from P0, P2, P5, and P10 satellite cells. Green indicated MyHC; blue indicated DAPI. Scale bars, 50 μm. (g) Quantification of MyHC⁺ cell numbers differentiated from satellite cells being serially expanded in vitro. Error bars represented S.E.M. and were based on three independent experiments. Significance was analyzed by student’s t-test. * indicated P<0.05, ** indicated P<0.01, *** indicated P<0.001. NS indicated no significant difference.