Buffalo (Bubalus bubalis) Fetal Skin Derived Fibroblast Cells Exhibit Characteristics of Stem Cells

Culture and characterization of fetal derived cells have received particular attention because of their easy access and possible source of stem cells for the study of development and differentiation. The present study was carried out to establish buffalo fetal fibroblast cell culture, and their longevity, expression patterns of pluripotency markers with prolonged passage and in vitro induced differentiation ability. Buffalo fetal fibroblasts were isolated from sub-dermal region and their primary culture was initiated in re-calcified buffalo plasma drops. On sufficient growth of primary culture, these cells were trypsinized and passaged at 80% confluency with a split ratio of 1:2 for multiplication of cells. Cryopreservation of cells was also performed at intervals of passages (P) 5 from confluent cultures, and the representative cells were allowed to proliferate in continuous cultures. These cells started emerging and anchoring to cell culture flasks within 24 h and survived up to P47 in 185 days with average passage time of 3.9 days. Expression of alkaline phosphatase and pluripotency genes viz., OCT-4, NANOG and SOX-2 were examined up to P45. Further, changes in relative expression of transcriptional factors were determined by quantitative real time PCR and found up-regulation of all the three genes up to P15 followed by up-regulation of SOX-2 up to P45 but down-regulation of Nanog. Upon induced differentiation, these cells differentiated into adipogenic and osteogenic cells as confirmed by oil red O and alizarin red stains, respectively. This study indicates that buffalo fetal fibroblast cells have characteristics of stem cells.