In vitro supplementation with the porcine plasma product, betaGRO?, stimulates activity of porcine fetal myoblasts and neonatal satellite cells in a divergent manner

摘要

Two separate experiments were conducted to evaluate the effect of betaGROsup?/sup supplementation on in vitro porcine fetal myoblasts (PFM) and porcine satellite cells (PSC) proliferation, fusion and myotube thickness. The PFM and PSC were isolated from the m. longissimus dorsi of day 60 of gestation fetuses and piglets within 24 h of birth, respectively. Proliferation assays were conducted as 4×3 factorial arrangements with time of culture (24, 48, 72, 96 h) and media treatment (standard porcine media supplemented with 10% (vol/vol) fetal bovine serum (HS); HS without 10% fetal bovine serum (LS); and LS supplemented with 10 mg/ml betaGROsup?/sup (BG)) as main effects. Fusion and myotube growth assays were conducted as 2×2 factorial designs with serum concentration (HS or LS), and betaGROsup?/sup inclusion (0 or 10 mg/ml) as main effects. There was a treatment×time interaction and betaGROsup?/sup×serum interactions for proliferation, fusion and myotube thickness of PFM (P 0.01). At all-time points, HS and BG-PFM had greater proliferation rates compared LS (P 0.01). The HS treatment had greater proliferation rates than BG (P 0.02) except at 72 h of culture (P=0.44). When betaGROsup?/sup was added to LS media, fusion percentage and myotube thickness decreased (P 0.01), while fusion percentage increased (P 0.01) and myotube thickness was unaffected (P=0.63) when betaGROsup?/sup was added to HS media. There were treatment×time and betaGROsup?/sup×serum interactions for proliferation rate and fusion rate of PSC, respectively (P 0.01). At all-time points, HS had greater proliferation rates than LS and BG (P 0.01), and LS had greater proliferation rates than BG (P 0.02). When betaGROsup?/sup was added to LS and HS media, fusion percentage increased for both media types (P 0.01). There was no betaGROsup?/sup×serum interaction (P=0.63) for PSC myotube thickness; however, betaGROsup?/sup supplemented myotubes were thicker (P 0.01) than non-betaGROsup?/sup supplemented myotubes. These two experiments indicate in vitro betaGROsup?/sup supplementation stimulates divergent responses based on the age of cell examined.