An Alternative Method for Long-Term Culture of Chicken Embryonic Stem Cell In Vitro

Figure 5

In vitro and in vivo differentiation of cultured cESCs. (a) Embryoid body-like structures in the phase after the cESCs were cultured without the growth factors for 10 days. (b) Oil Red O positive staining showed the differentiation to adipocytes in the phase. cESCs with complete culture medium in the phase were the control. (c) The expression of PPARγ (red) confirmed the differentiation to adipocytes, and the nucleus was stained by DAPI (blue). (d) Alizarin Red staining verified the potentiality to osteoblasts. cESCs with complete culture medium in the phase were the control. (e) The immunofluroscence of desmin (red) and DAPI (blue) certified the directional differentiation to smooth muscle cells. (f) MHC (red) and DAPI (blue) staining proved differentiation to cardiac myocytes. (g) The immunofluorescence of PAX6 (green) and DAPI (blue) showed the differentiation to neural cells. (h) The process of chimeric chicken production was indicated in the figure. Chimeric chickens generated after grafting cultured cESCs of 25 passages in vitro. (i) The two distinct genotypes of PRLpro2 on the gel picture verified that there were two successful productions of chimeric embryos. PCR analysis of PRLpro2 genotype. Six chicken embryos after 15 d, 12 d, 10 d, 8 d, and 13 d incubation were tested. Scale bar: 100 μm.

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