Figure 3
From: Phenotypic differences in hiPSC NPCs derived from patients with schizophrenia
Aberrant migration in schizophrenia (SZ) human-induced pluripotent stem cell (hiPSC) forebrain neural progenitor cells (NPCs). (a) Representative images of hiPSC forebrain NPC neurosphere outgrowth assay. The average distance between the radius of the inner neurosphere (dense aggregate of nuclei) and outer circumference of cells (white dashed line) was calculated. 4′,6-Diamidino-2-phenylindole (DAPI)-stained nuclei (blue). Scale bar, 100 μm. (b) Neurosphere outgrowth by control and SZ hiPSC forebrain NPCs. (c) Neurosphere outgrowth following coculture with control or SZ hiPSC NPC-conditioned media (CM). (d) Representative images of mixed control and SZ hiPSC NPCs, labeled with lentivirus-green fluorescent protein and lentivirus-red fluorescent protein. Green and red numbers indicate five furthest migrated lentiviral-green fluorescent protein and lentiviral-red fluorescent protein NPCs in reciprocal migration experiments. DAPI-stained nuclei (blue). Scale bar 100 μm. (e) Neurosphere outgrowth in neurospheres composed of mixed control and SZ hiPSC NPCs. (f) Representative images of hiPSC forebrain NPCs after 5 days in a microfluidic device. At time 0, all NPCs were below the microgrooves pictured—migration occurred up from chamber B into chamber A. Migrating neural cells stained with MAP2AB (red), βIII-tubulin (green); DAPI-stained nuclei (blue). Scale bar, 100 μm. (g) Schematic of the microfluidic chambers. At time 0, hiPSC NPCs were added to chamber B and allowed to begin migration toward chamber A via 15-μm grooves. (h) Cellular migration of control and SZ hiPSC NPCs in microfluidic devices at 48 h. (i) Representative images of hiPSC forebrain NPCs in micropatterned laminin spot migration assay. Laminin spot stained (green); migrating neural cells stained with βIII-tubulin (red); and DAPI-stained nuclei (blue). Scale bar, 100 μm. (j) Nearest-neighbor analysis of ‘chaining’ between laminin spots by control and SZ hiPSC forebrain NPCs. Error bars are s.e.m., *P<0.05, **P<0.01, ***P<0.001. See also Supplementary Figures 6–8.
