Isolation and characterization of buffalo amniotic fluid stem cells

Abstract

Amniotic fluid (AF) present different cellular types in its composition and a specific group of these cells shows multipotent characteristics with high proliferation ratio and increased differentiation ability. In this study, we isolated and characterized adult stem cells from buffalo (Bubalus bubalis) to check their differentiation potential in-vitro and evaluated their plasticity. Single cell derived stem cell clones were isolated by limiting dilution. AF cells showed both fibroblast and epitheloid morphology. Amniotic fluid stem cells (AFSC) were differentiated into osteogenic, chondrogenic and adipogenic lineage by culturing in specific differentiation medium. Differentiation was confirmed by cell staining with alizarin red S, toluidine blue and oil red O specific for osteogenesis, chondrogenesis and adipogenesis respectively. Gene expression of stem cell markers were found to be positive for Oct- 4, Sox–2 and Nanog by RT- PCR.