Phenotypic differences in matrix metalloproteinase 2 activity between fibroblasts from 3 bovine muscles

Abstract

Different muscles in a beef carcass are known to respond differently to the same stimulus or animal growth pattern or both. This may complicate the search by the meat industry for production methods to render meat tender. One of the major differences between muscles in the same carcass is in the expression of intramuscular connective tissue. Current study investigates the existence of a phenotypic difference among fibroblasts from 3 bovine skeletal muscles as exemplified by the expression of matrix metalloproteinases (MMP) the main enzymes responsible for connective tissue turnover. The sensitivity of phenotypic differences to cell culture conditions (passage number, presence of growth factors from fetal serum) was also examined. Fibroblasts, the main cells responsible for the production and turnover of collagen were isolated from LM, semitendinosus (STN), and sternomandibularis (SMD) muscles from a bull calf and grown in DMEM, 10% fetal bovine serum, and 5% CO(2). Cell doubling times, survival time, resting expression, and activity of MMP and the effect of serum withdrawal in the culture media on MMP expression and activity were determined for each cell line during 15 passages. Fibroblasts isolated from the 3 muscles had different growth potentials. The shortest (P < 0.0001) cell doubling times for almost every passage were found in cells from STN muscle. Cells from the LM had a shorter (P < 0.0001) survival time in comparison with STN and SMD. Cells derived from the STN had greater values (P > 0.05) of MMP-2 activity in comparison with LM and SMD cells until passage 4. At passage 15, no activity was detected for any cell line. Serum withdrawal generally reduced MMP-2 activation but did not eliminate differences in activity between fibroblasts from the 3 muscles. These results suggest that fibroblasts from different locations are phenotypically different and may respond differently to the same growth or nutritional stimulus in vitro. This may be related to in vivo differences in accumulation, maturity, and turnover of collagen, and ultimately meat tenderness. These findings may be important for selecting a management strategy for improving meat tenderness by manipulation of animal growth; a strategy applied to the whole animal may work for some muscles but not for others.