We constructed an hESC line with Citrine (Cit) fused to the endogenous copy of the DCX gene in the H1 parent line to allow us to profile neurons (Cit+) and progenitors (Cit−) (Yao, 2017). A majority of cells were FOXG1 (76.2 ± 2.9%) and NKX2-1+ (85.7± 0.3%) by day 24 (Figure 1J, N), indicating that the DCX-citrine line differentiated comparably to the parent line (Figure 1N; compare with Figure 1F). By D54, we observed many GAD67+Cit+ cells (Figure 1K), but no overlap between MKI67 expression and Cit (Figure 1L). FACs analysis indicated that the percentage of Cit+ cells in these cultures increased between D19 and D54, declining thereafter (Figure 1M). At D54, 89 ± 2.3% of Cit+ cells were MAP2+, indicating Cit was confined to post-mitotic neurons (Figure 1O). In addition, 80.5 ± 6.8 % of Cit+ cells were GAD67+, and 32.6 ± 3.5% were SST+ (Figure 1O), indicating the DCX-Cit line will allow us to enrich for GABAergic cells for transcriptome profiling.
