INSULIN-LIKE GROWTH FACTOR (IGF)-I AND -II AND IGFBP SECRETION BY OVINE SATELLITE CELL STRAINS GROWN ALONE OR IN COCULTURE WITH 3T3-L1 PREADIPOCYTES

摘要

The current study was designed to examine the effects of muscle and fat stem cell coculture on the secretion of insulinlike growth factor (IGF)-I and -If and IGF binding proteins (IGFBP) by these cells. Two sheep satellite cell strains with negligibleor high potential for differentiation (10A and 0_1, respectively) were placed in coculture with 3T3-L1 preadipocytes using a filter support to separate the two cell types. Media conditioned by the cells grown alone or in coculture were analyzed for IGFsby RIA or IGFBPs by ligand blotting. The numbers of satellite cells and preadipocytes declined throughout the 5-d culture period, although coculture slowed the 3T3-L1 decline but hastened the satellite cell decline. The satellite cell strains and 3T3-L1cells secreted small amounts of IGF-I (<= 2 ng/ml) and IGF-II (< 10 ng/ml) over the 5-d culture period, Coculture did not increase the amount of IGF-I and -II in conditioned media. The lowly differentiating 10A cells secreted barely detectable amounts of the low molecular weight IGFBP-3 subunit (34 kDa), IGFBP-2 (28 kDa), and IGFBP-4 (18 kDa). Coculture of 10A and 3T3-L1 cells potentiated secretion of IGFBP-2 and -3. Strain 0_1, which readily differentiates, secreted high levels of both IGFBP-3 subunits (34 and 39 kDa) and IGFBP-2 (28 kDa), as well as significant amounts of the 18 kDa IGFBP-4, Coculture did not alter IGFBP secretion of 0_1 cells. This study showed that while IGF-I and -II levels in media conditioned by sheep satellite cell strains arelow and relatively invariant, the intensity and complexity of IGFBP patterns increases with time in culture and with the potential for differentiation of the satellite cell strains. Coculture with preadipocytes appeared to potentiate IGFBP secretion while reducing satellite cell viability.