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.
KEY WORDS:Cell culture granulosa-luteal hypotonic lysis IVF cells