Fig. 4: Chicken ES cells can contribute to chimera formation both ex ovo and in ovo.
From: Derivation of embryonic stem cells across avian species
a, GFP-labeled chicken ES cells were implanted onto ex ovo cultured EGK.X or HH4 chicken embryos. Scale bars, 100 μm (cES cells), and 400 μm (EGK.X and HH4 embryos). b, Representative images showing HH4-HH11/12 stage embryos developed from ex ovo cultured chicken EGK.X embryos without the implantation of chicken ES cells. Scale bars, 400 μm. c, Representative images showing HH4-HH11/12 stage embryos developed from ex ovo cultured chicken EGK.X embryos implanted with GFP-positive chicken ES cells. Scale bars, 400 μm. d, Representative images showing HH8/9 and HH11/12 stage embryos developed from ex ovo cultured chicken HH4 embryos implanted with GFP-positive chicken ES cells. Scale bars, 400 μm. e, Image of the surrogate open-shell system for in ovo chimeric embryo development. f,g, Chimeras generated by injecting GFP-positive chicken ES cells into EGK.X-stage chicken embryos: medium-grade E5 chimera (f) and high-grade E4 chimera (g). Scale bars, 400 μm. EXE, extra-embryonic blood vessels (scale bar, 100 μm). h, Images showing control, low-grade, medium-grade and high-grade E20 chimeras generated by injecting RIR chicken ES cells into EGK.X White Leghorn chicken embryos. Scale bar, 10 mm. i, A chimeric chicken with black feathers from donor ES cells derived from RIR at 31 days old posthatch. Recipient embryo, White Leghorn. j, Images showing GFP-positive cells in the indicated organs of chimeras generated by injecting GFP-positive chicken ES cells into EGK.X chicken embryos. Heart (E13, scale bar, 1 mm), tongue (E20, scale bar, 1 mm), midbrain (E8, scale bar, 100 μm), stomach (E7, scale bar, 400 μm), liver (E21, scale bar, 400 μm) and intestine (E19, scale bar, 400 μm). Chimeras generated are from 1 representative experiment of more than 5 experiments performed in a–d, for ex ovo, and more than 100 experiments performed in e–j, for in ovo.
