Isolation, in-vitro cultureand characterization of primordial germ cells of Kadaknath breed of chicken

The aim of this study was to harvest the primordial germ cells (PGCs) sourced from circulating-blood of early embryos from Kadaknath. Considering PGCs as the precursors of sex cells, which are documentedto be the best route to generate a germline chimera, compared to other early embryonic cells, the same needed to be delineated in Kadaknath chickens, for their prospective uses in future. Fresh Kadaknath fertile-eggs were incubated for 50 h to reach the embryonic H&H stages: 11–17. The circulating blood,1–5 µl was collected from the Kadaknath embryos blood vessels through skilled hands and cultured in-vitro following standard protocols. The purified PGCs were then characterised using Periodic Acid Schiff’s, Oil Red O and Stage Specific Embryonic Antigen–1, which successfully demonstrated their conformities to the pluripotent marker standards. The study concluded that, circulating PGCs of Kadaknath native chickens, could be efficiently harvested and propagated for their possible uses as bio-banking potent cells, in future’s avian germplasm conservation program. Therefore, this research work registers the first successful isolation and in-vitro culture of blood-derived Kadaknath PGCs and their characterization.