ONTARIO, CANADA

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.

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