A simplified method for preparing IVF granulosa cells for culture

PURPOSE: To validate an expedited method for the removal of erythrocytes when preparing IVF granulosa-luteal cells for culture. METHODS: Granulosa cells were enriched for culturing from follicular aspirates by density gradient centrifugation and by hypo-osmotic lysis treatments. RESULTS: Cells prepared by either method showed the same cell viability and produced progesterone in similar quantities. CONCLUSIONS: Using hypo-osmotic lysis to remove erythrocytes does not impair granulosa cell viability or steroidogenesis. It avoids multiple density gradient centrifugations and washings, and yields IVF granulosa cells ready for culture efficiently.