Asian Journal of Animal and Veterinary Advances1683-99191996-3289Academic Journals Inc.10.3923/ajava.2011.772.788HepsibhaP. MeenambigaiT.V. MangalagowriA. PalanisamyA. StalinA. NithyaS. KumananK. 8201168The present study has been undertaken for isolation, characterization and differentiation of Buffalo Adipose Derived Mesenchymal Stem Cells (bADMSCs). Cocktail enzymatic digestion method was used for isolation of bADMSCs which yielded optimum number of cells than other methods. Subcutaneous adipose tissue yielded comparatively higher number of Stem Cells (SC) than from omentum and bone regions. The growth and proliferation of SC were rapid when Mesencult medium was used, in comparison to Dulbeccos Modified Eagles Medium (DMEM). Expression of oct 4 gene by reverse transcriptase-polymerase chain reaction (RT-PCR) indicated the stemness of isolated bADMSCs. Magnetic cell sorting and immunostaining revealed that the isolated ADMSCs were cluster of differentiation 44 (CD44) positive. 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