The present study was carried out with the aim to isolate, culture and characterize the totipotent stem cells from in vitro derived embryos and pluripotent stem cells from a month-old fetus in buffalo. The oocytes were matured, fertilized and the embryo development upto 16 cell stage were recorded under 15% O_2, 5% CO₂, and 38.5 ºC temperature and 90% humidity. The overall cleavage rate and morula development was 9.1 % and 4.6% respectively. The results indicated that blastomerers from 2 cell, 4 cell and 8 cell state embryo did not grow and degenrated. However, blastomere cells isolated from 16 cell stage embryos cleaved and formed clones. Pluripotent stem cells were derived from 30-40 days old buffalo fetus and cultured on buffalo fibroblast cell monolayer with RPMI-1640 and 15% FBS in the presence of LIF. The pleuripotent stem cells and totipotent s were characterized and found positive for SSEA-1,4, EMA1 and alkaline phosphatase and negative for SSEA-3. From this result it was concluded that totipotent and pleuripotent stem cells shall be identified differently and more specific markers needed to study before using these cells for specific reproductive technologies

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P V Mehere

Assistant Professor, Department of Veterinary Physiology KNP College of Veterinary Science,

Shirwal-412801, Maharashtra, India