Ovarian Granulosa Cells Isolated with EGTA and Hypertonic Sucrose: Cellular Integrity and Function

Abstract

Chemical treatments previously shown to disrupt gap junctions were applied to rat ovaries with antral follicles prior to expressing the granulosa cells with a blunt spatula. Exposure of the ovaries to 6.8 mM EGTA [ethyleneglycol-bis-β-aminoethyl ether)-N,N'-tetracetic acid] and 0.5 M sucrose either by perfusion in situ or incubation in vitro generated monodisperse suspensions of granulosa cells with improved integrity as evaluated by several criteria.