A simplified method for preparing IVF granulosa cells for culture.
OBJECTIVE
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.
Department of Obstetrics and Gynecology, McMaster University, Room No. 3N52 Health Science Centre, 1200 Main St. W., L8N 3Z5, Hamilton, Ontario, Canada. lobbd@mcmaster.ca
Derek K
DK
Lobb
Edward V
EV
Younglai
D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000166:cytology; D006107:Granulosa Cells / Q000166:cytology*; D006801:Humans