Figure 1.

Induction of floor plate progenitors from hESCs at day 11 using a new developmental-based protocol

(A) Schematic of the differentiation protocol for mDA neurons.

(B) Violin plots of LMX1A, FOXA2, and OTX2 generated from scRNA-seq data of developing human ventral midbrain shown across corresponding cell types. Right axis shows absolute molecular counts. Gray, enriched over baseline with posterior probability >99.8%. For cell type nomenclature, see La Manno et al. (2016).

(C) Gene expression profile of differentiated cells according to CHIR99021 concentration at day 11.

(D and E) Immunostaining of LMX1A⁺;FOXA2⁺ cells (D) and LMX1A⁺;OTX2⁺ cells (E) at day 11. Scale bar, 200 μm.

(F) Quantification of LMX1A⁺;FOXA2⁺ cells and LMX1A⁺;OTX2⁺ cells. NS, not significant (n = 4 independent experiments).

(G) qPCR analysis of GBX2 and HOXA2 in cells differentiating on either LN111 or LN511. ^∗p < 0.05 versus D0; †p < 0.05 versus LN511 (n = 6 independent experiments).

(H) Cell yield by LN111 or LN511 at day 11. ^∗∗p < 0.01, ^∗∗∗p < 0.001 versus D0; †p < 0.05 versus LN511 (n = 6 independent experiments).

(I) qPCR analysis of differentiated cells in the presence/absence of WNT5A at day11 (n = 5–9 independent experiments). ^∗p < 0.05 versus WNT5A(−) condition.

(J and K) Immunostaining of LMX1A⁺; FOXA2⁺ cells (J) and LMX1A⁺;OTX2⁺ cells (K) at day 11. Scale bar, 200 μm.

(L) Quantification of LMX1A⁺;FOXA2⁺ cells and LMX1A⁺;OTX2⁺ cells. ^∗∗p < 0.01, ^∗∗∗p < 0.001 versus WNT5A(−) condition (n = 4 independent experiments).