In Vitro Characterization of Motor Neurons and Purkinje Cells Differentiated from Induced Pluripotent Stem Cells Generated from Patients with Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay

Figure 1

Characterization by immunofluorescence of MNs and Purkinje cells differentiated from healthy or ARSACS iPSCs cultured in 2D. MNs and Purkinje cells differentiated from iPSCs were seeded on poly-D-lysine coated wells and differentiated until days 13 and 14, respectively. MNs were characterized by immunofluorescent staining using the CHAT (a, b) (in green) and Islet-1 (c, d) (in green) specific MN markers and the neuronal markers NFM (c, d) (in white) and β3-tubulin (a–d) (in red). Purkinje cells were stained with neuronal markers β3-tubulin (e–l) (in red) and NFM (k, l) (in white) and with Purkinje cell-specific markers GRID2 (e, f) (in green), KIRREL2 (g, h) (in green), parvalbumin (i, j) (in green), PCP2 (k, l) (in green), and calbindin (k, l) (in red). Purity was assessed by immunofluorescence and counting MNs stained with Islet and β3-tubulin (m), and Purkinje cells were stained with KIRREL2 and β3-tubulin (n) (see raw data in the table). Nuclei in (a)–(d) and (k) and (l) were stained in blue with DAPI. Scale bar in (l): 20 μM.