Generation and characterization of induced pluripotent stem cells in domestic Asian water buffalo (Bubalus bubalis)

Induced pluripotent stem cells (iPSCs) have numerous applications in livestock to improve production traits, disease resistance, biopharming, conservation of germplasm, disease modeling, regenerative medicine etc. Here, we have described the derivation of iPSCs from buffalo fetal fibroblasts (BuiPSCs) by lentivirus based transduction of mouse derived pluripotency marker genes Oct4, Sox2, Klf4 and c-Myc. The BuiPSCs showed typical buffalo embryonic stem cells like colony morphology which were alkaline phosphatase (AP) positive and expressed pluripotency markers Oct4, Nanog, SOX2, KLF4, FoxD3 and SSEA1, TRA-1-60, TRA-1-81. The cells were carrying normal karyotype and were able to differentiate into cell of all three germ layers in vitro. The BuiPSCs could be propagated beyond 20th passages. To the best of our knowledge, this is the first report on generation of buiPSCs in domestic water buffalo (Bubalus bubalis) using mouse derived transcription factors and the reprogrammed cells could self renew more than 20th passage