Reprogramming of buffalo (Bubalus bubalis) foetal fibroblasts with avian egg extract for generation of pluripotent stem cells

An attempt was made to study the efficiency of chicken (Gallus gallus domesticus) egg extract (EE) to reprogram buffalo (Bubalus bubalis) foetal fibroblasts (bFFs) without incorporation of ectopic transcription factors. The isolated bFFs were cultured in media supplemented with 2%, 4%, 6% and 10% EE to induce reprogramming. It was observed that fewer but larger sized alkaline phosphatase positive (AP+ve) colonies developed in culture system containing 2% EE whereas, more but smaller sized colonies developed in 4%, 6% and 10% EE. The developed colonies expressed pluripotency markers like Oct4, Nanog, SSEA1, TRA-1-60, TRA-1-81 and RT-PCR study revealed relative expression of genes indicating pluripotency (Oct4, Sox2, Nanog and FoxD3) increased as the concentration of EE increased in culture systems confirming the reprogramming capability of chicken EE.