Extracellular matrix associated with human luteininzing granulosa cells

Abstract

Human granulosa cells (GC), prepared from follicular aspirates using a non-enzymic method, were maintained on chamber slides in a defined medium without additional attachment factors. In this system, GC clustered to a limited extend and attached only loosely to the substream necessitating medium replacement through repeated partial changes to avoid cell loss. Cell size and progesterone production increased consistently with continuing luteinization. These processes were associated with maintenance and deposition of the endogenous extracellular matrix components heparan sulphate proteoglycan and laminin (α2, β1, γ1 subunits) as shown by both Western blotting and immunohistochemistry. Collagen IV seemed to be present between freshly prepared GC and was maintained in culture. The addition of either epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) into the culture medium did not reveal any marked effect on deposition of laminin components during culture. Release of cell clusters, particularly in the absence of gonadotrophin, was noted during the later stages of culture, constituting the so-called 'lift off' phenomenon. This has been correlated with an increasing concentration of matrix metalloproteinases (MMP-2) in the culture medium during the culture period. To conclude, luteinization and maintenance of the GC-derived layer of the corpus luteum is likely to involve deposition and conservation of pericellular extracellular matrix components, actively synthesized by the GC themselves and controlled by the presence of gonadotrophin.

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