Figure 3.

In vitro assays for human NSPCs

(A) Typical sort layout for limiting dilution assays to measure neurosphere initiation frequency.

(B) Representative images of neurospheres, indicative of a positive well. Scale bar 100 μm.

(C) Immunofluorescence (IF) images of CD24⁻THY1^−/lo populations differentiated in vitro for 5 days, then stained for GFAP (red), DCX (green), O4 (magenta), or DAPI (blue).

(D) IF images of THY1^hi populations differentiated in vitro for 5 days, then stained for GFAP (red), O4 (green), or DAPI (blue). Abbreviations: E, EGFR; P, PDGFRA.

(E) IF images of CD24⁺THY1^−/lo populations differentiated in vitro for 5 days, then stained for DCX (green) or MAP2 (red). Scale bar 50 μm. Adapted from Liu et al., 2023.¹