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In vitro Growth of Muscle Satellite Cells Isolated from Normal and Callipyge LambsRodriguez, Beatriz T. 01 May 1999 (has links)
The muscle hypertrophy of lambs expressing the Callipyge phenotype is possibly linked to characteristics of their muscle satellite cells. Therefore, characteristics (proliferation, fusion %, and protein accretion) of cultured satellite cells isolated from the longissimus muscle of Callipyge (n = 3) and normal (n = 3) lambs were compared in this study. In the first experiment, we tested whether or not the lll proliferation rates differ for satellite cells isolated from Callipyge or normal sheep when cultured in the presence of different serum types (horse, normal lamb, or Callipyge lamb). The average population doubling time (PDT, h) during log phase growth was calculated for cells from each animal grown in each serum type. Population doubling time was not affected (P > .1) by the interaction of satellite cell type with serum type, or by satellite cell type. Unexpectedly, PDT was longer (P < .05) for satellite cells grown in Callipyge serum (22 h) than for cells grown in normal sheep serum (20 h) or horse serum (18 h). These results suggest that muscle hypertrophy of Callipyge lambs is not linked to intrinsic differences in satellite cell proliferation, although hypertrophy may be associated with a decreased proliferation induced by a factor in Callipyge serum.
In the second experiment, we tested whether cell fusion, or protein accretion differ for cultured satellite cells isolated from Callipyge or normal sheep. DNA and protein were determined at 24, 48, 72, and 96 h after satellite cell cultures were induced to differentiate. Fusion percentage was determined in a Giemsa stained plate after 72 h in differentiation medium (Dulbecco's Modified Eagle Medium containing 1.5% of horse serum). Callipyge cultures tended (P = .14) to have higher fusion% than normal cultures exhibited, suggesting that muscle hypertrophy of Callipyge lambs may be linked to an increased tendency of satellite cells to fuse. Protein content (μg/well) and protein:DNA ratio (ng of protein/ng of DNA) were not affected by satellite cell type (P = .80 and P = .79, respectively). Thus, there was no evidence for a link between increased protein accretion and Callipyge hypertrophy.
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