77 Determination of Cellutein and Betagro Mode of Action within Porcine Satellite Cell Cultures

Abstract

Abstract The objective of the study was to evaluate the efficacy of celluTEIN and betaGRO to influence porcine myogenesis in vitro. Satellite cells (N = 3 piglets) were plated on ECL-coated 12-well plates at a density of 5,000 cells/cm2. Cells were grown to confluence in growth media containing 10% fetal bovine serum, 100 U penicillin/mL, 100 µg of strepmycin/mL, and 20 µg of gentamicin/mL in high glucose DMEM for 72 hours. Cells were treated with 1 of 4 treatments: 2% fetal bovine serum, 100 U penicillan/mL, 100 µg of strepmycin/mL, and 20 µg of gentamicin/mL (CON), CON+5 ng/mL recombinant IGF-1 (IGF-1), CON+10 mg/mL betaGRO (BG), CON+10 mg/mL celluTEIN (CT). For myotube width assays, FBS was reduced to 2% when cells were confluent for 96 hours, at which time treatments above were applied with or without 20 µM rapamycin. Fusion capacity (FC) and myotube width (MW) were determined from 5 representative photomicrographs and experiments were repeated 3×. There were treatment effects for FC and MW. Cells treated with CT and BG had greater FC and MW than the other treatments (( P < 0.05), but did not differ from each other (P > 0.05). Cells treated with IGF-1 had greater FC and MW than CON cells (( P < 0.05). There was a Treatment×Rapamycin interaction (( P < 0.05). Without rapamycin, CON myotubes had smaller diameters than other treatments (( P < 0.05), BG and CT myotubes were greater than IGF-1 (( P < 0.05), but were not different (P > 0.10) from each other. Though there was a statistical difference between treatments when rapamycin was added (P < 0.05), treatment differences did not reflect a difference that can be translated out of the satellite cell model. These data illustrate that celluTEIN and betaGRO stimulate porcine satellite cell differentiation and growth through mechanistic target of rapamycin signaling.