Derivation and Expansion Using Only Small Molecules of Human Neural Progenitors for Neurodegenerative Disease Modeling (original) (raw)
Figure 3
Differentiation of PNS neurons and mesenchymal cells from neural epithelial cells.
(A) Summary of isolation and differentiation protocol used in this study. (B) Immunostaining of differentiated neural epithelial cells for HNK-1. (C) Neural epithelial cells were treated with CHIR for 2 days and then switched to BMP4 or serum-containing medium for 2 additional days. In both cases, the cultures show cells positive for the neural plate border/neural crest markers PAX7 and SLUG, whereas only some cells are still positive for PAX6. (D) Immunostaining of smNPCs differentiating in the presence of BMP4 for PERIPHERIN and BRN3A. (E) qRT-PCR demonstrating the upregulation of PERIPHERIN and BRN3A in neural epithelial cells differentiated for 8 days in the presence of BMP4, but not PMA, following two weeks of maturation. (F) More than 40% of cells are double positive for PERIPHERIN and TUBBIII after patterning with BMP4 and maturation. Error bars represent variation from 2 independent cultures. Scale bars are 100 µm. CHIR = CHIR99021, DM = dorsomorphin, FCS = fetal calf serum, PMA = purmorphamine, and SB = SB43152.