Differentiation of iPSCs into NPCs was initiated, as previously described (Maguire et al, 2016 (link)) with indicated modification. Briefly, cultures were treated with daily media changes containing SB431542 (10 μM; Tocris), LDN193189 (1 μM, Tocris), and endo‐IWR1 (1.5 μM; Tocris) and supplemented with B27 without vitamin A (Invitrogen) and passaged at days 4 and 8 of differentiation. From days 8 to 14 of differentiation, NPCs were expanded in Invitrogen neural expansion media, containing Neural Induction Supplement in Advanced DMEM/F12 and Neurobasal Medium (Invitrogen, per manufacturer instructions). On Day 14, NPCs were cryopreserved after confirmation of NPC identity with > 90% expression of Forse‐1.
Schlotawa L., Tyka K., Kettwig M., Ahrens‐Nicklas R.C., Baud M., Berulava T., Brunetti‐Pierri N., Gagne A., Herbst Z.M., Maguire J.A., Monfregola J., Pena T., Radhakrishnan K., Schröder S., Waxman E.A., Ballabio A., Dierks T., Fischer A., French D.L., Gelb M.H, & Gärtner J. (2023). Drug screening identifies tazarotene and bexarotene as therapeutic agents in multiple sulfatase deficiency. EMBO Molecular Medicine, 15(3), e14837.
Endo Ipscs Ldn193189 Neural Neural expansion Sb431542 SupplementVitamin a
Corresponding Organization :
Other organizations :
Children's Hospital of Philadelphia, University of Southampton, German Center for Neurodegenerative Diseases, Telethon Institute Of Genetics And Medicine, University of Naples Federico II, University of Washington, Bielefeld University, Baylor College of Medicine, Neurological Research Institute, Universitätsmedizin Göttingen, University of Göttingen
Expanded in Invitrogen neural expansion media from days 8 to 14
Supplemented with B27 without vitamin A
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
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