![]() |
Figure 4: The giPSCs were differentiated into germ cells like cells. (a) Embryoid bodies cultured in the presence of Retinoic acid (RA) and bone morphogenetic protein 4 (BMP4) shows different germ cells characteristics. (b) Vasa, Dazl, Stella, Scp3, Pum8, Nanos1 and Nanog genes expression at transcription level in germ cells like cells generated from giPSCs after 50 days of differentiation, highlights the basic germ cell characters. Gapdh is a housekeeping gene and Nanog has been taken as positive marker in goat induced pluripotent stem cells (giPSCs) and germ cell-like cells (giPS-GC) generated from giPSCs. (c) Presence of Vasa, Dazl, Scp3 and Stella protein in immune-staining confirms the presence of germ cells characters in differentiated giPSCs. (One scale bar is 200 μm). (d) Vasa, Dazl and Scp3 are not observed after 10 days of differentiation while Stella is expressed by differentiated germ cells like cells. Expression of Vasa, Dazl, Stella, and Scp3 increases after 20 days of differentiation. (e) Presence of Vasa protein (light band) in western blotting assay of giPS-GC lysate affirms germ cell differentiation. (f) Fluorescence-activated cell sorting (FACS) assay showed the 0.1 % germ cells like cells in differentiated population of giPSCs after 10 days of differentiation. |