Figure 6.

FACS for GD2 Enriches SAP-like Cells

(A) qPCR analysis of SA genes in p75NTR+/GD2+, p75NTR+/GD2− H9 NCPC-4d, and NCPC-6d cells. Expressions were normalized to that of hESCs. ND, not detectable at <35 cycles. N = 3 (4d) and N = 3 (6d) independent experiments.

(B) NCPC-4d p75NTR+/GD2+ and p75NTR+/GD2− cells after 6 days differentiation in DP express SA (αTH), chromaffin (PNMT), and neuronal (PRPH) markers. Scale bar, 5 μm. After DP differentiation, PNMT and PRPH are extensively co-expressed, and proportions of these cells are both increased in the GD2+ population, and all αTH+/GD2+ cells expressed PNMT. DAPI was used to count the total cells. N = 4 independent experiments.

(C) qPCR analysis of NCPC-4d p75NTR+/GD2+ cells differentiated with DP for 6 and 9 days. GD2 preselection augments PNMT expression as well as other SAP genes. ND, not detectable, N = 4 independent experiments.

Error bars represent mean ± SEM. ns, not significant, ^∗p > 0.05, ^∗∗p > 0.01, ^∗∗∗p > 0.001, ^∗∗∗∗p > 0.0001.