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 2

Immunofluorescence characterization of MNs and Purkinje cells differentiated from ARSACS iPSCs cultured in 3D. ARSACS iPSC-differentiated Purkinje cells were cultured on sponges containing fibroblasts for 53 days (a–d). ARSACS iPSC-differentiated MNs were cultured on sponges populated with fibroblasts and ARSACS iPSC-derived Schwann cells for 74 days (e, f). The 3D models were imaged by immunofluorescence through a view from above after staining of cells with the neuronal marker NFM (c, d) in green and with the specific markers calbindin (a), KIRREL2 (b), and parvalbumin (c) in white, PCP2 (d) in red for Purkinje cells, NFM in green, ChAT (e, f) in red, and Islet1 (f) in white for MNs. These results are representative of the three ARSACS cell populations. Nuclei were stained in blue with DAPI. Scale bar in (f): 20 μm.