Expression of the IGF system in primary and immortalized porcine ovarian granulosa cells

We have established a novel granulosa cell line derived from porcine ovarian follicles (4-6 mm in diameter). This cell line, MDG2.1, was obtained by transfection of freshly cultured cells with the plasmid pSV3neo. Doubling time for MDG2.1 cells is 24-36 h. Northern analysis for RNAs of the insulin-like growth factor (IGF) regulatory system indicates that RNAs for IGF-I, IGF-I receptor, and IGF binding proteins (IGFBP) 2-6, but not IGFBP-1, are expressed in MDG2.1 cells. The secretion of IGFBPs from MDG2.1 cells shows > 10-fold levels of the 24 kDa form, reduced secretion of other IGFBPs, with no change in the total amount of IGFBP secreted, as compared to primary cells cultured under identical conditions. Use of endoglycosidase F indicated that several IGFBPs are posttranslationally modified. This cell line is a useful model and plasmid transfection target system to investigate IGFBP action in ovarian granulosa cells.