Author : Sesha Subramanian, Aarti Raj, Rajesh Kumar, Satish Kumar Rana, Anal Kant Jha and Sanjeev Gautam

The current study was carried out to isolate, culture, characterize and cryopreserve the granulosa stem cells from buffalo ovaries. Granulosa stem cells were cultured in DMEM containing15% FBS, 1% penicillin/ streptomycin and 1%L-glutamine in 5% CO2 with humidified air at 38.5±0.5˚C. After 3 days of culture different morphologies of cells were observed. The confluent monolayer should have been formed at the base of the droplet after 72 hrs of culture. The culture was harvested and cultured in a culture media. Followed by cryopreservation of the granulosa stem cells. This result may contribute towards establishing further studies of granulosa stem cells for various therapeutic and reproductive biotechnological applications.

Keywords: Buffalo, granulosa stem cells, confluent monolayer, and characterization and reproductive biotechnology