Rearrangement of extracellular matrix during cluster formation by human luteinising granulosa cells in culture

Human ovarian granulosa cells were cultured on a basement membrane preparation (Matrigel) to investigate the role of extracellular matrix components in granulosa cell cluster formation. Time-lapse videomicroscopy of these cultures revealed a rapid aggregation of cells which was initiated during the first 2-4 h of culture so that by 8 h most of the granulosa cells were incorporated into clusters. Further amalgamation then occurred with the transfer of cells along 'bridges' between combining clusters. The clustering process, which was complete by about 24 h, was accompanied by reorganisation of matrix which was visualised by immunolabelling of laminin. Clustering cells appeared to gather matrix which became distributed around the clusters. Confocal microscopy showed matrix to be present over the surface of each cluster as well as around the base apparently anchoring the aggregate to the culture surface. Results suggest the potential for active rearrangement of matrix by granulosa-derived cells during corpus luteum development.