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. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Nat Protoc. 2019 Mar 25;14(4):1261–1279. doi: 10.1038/s41596-019-0141-y

Figure 4. Characterization of hPSC-derived ENC and enteric neurons.

Figure 4

a) Protocol for neuronal differentiation and maturation of ENC precursors, b) Flow cytometry analysis of CD49D positive ENC cells from hESC line UCSF4 and hiPSC line WTC11 on day 12. c) Flow cytometry analysis of CD49D positive ENC cells from hESC line UCSF4 and hiPSC line WTC11 after ENC spheroid enrichment on day 15. d) Immunofluorescence staining of TUJ1/TRKC on day 30 of EN induction. e) Flow cytometry analysis of TUJ1 and TRKC expression in EN cells on day 20, day 40 and day 55. f) Immunofluorescence images of CHAT, 5HT, NOS1, and GABA stained ENs on day 50. g) Flow cytometry analyses of CHAT, 5HT, NOS1, and GABA on ENS at day 75. AA, ascorbic acid; GDNF, Recombinant Human Glial Derived Neurotrophic Factor, F647, Alexa Fluor™ 647. Scale bars = 100 μm in c, f and 20 μm in e. o