Cls3261

Differentiation protocol was adapted from [16 (link)]. iPSC colonies were dissociated into small clusters (50–200 cells) with 0.5 mM EDTA (Sigma Aldrich, Darmstadt, Germany #E6758). The cell clusters were transferred with neural induction media made of 1:1 DMEM/F-12 Glutamax (Gibco #10565-018, Waltham, MA USA) media supplemented with N2 (Gibco #17502048) and Neurobasal media (Gibco #21103-049) supplemented with B27 (Gibco #17504044), 1% Penicillin/Streptomycin, 5 μg/mL Insulin (Sigma Aldrich #I6634), 20 ng/mL bFGF (PeproTech #100-18B, Rocky Hill, NJ, USA) and 20 ng/mL EGF (Sigma Aldrich #E9644) into an ultra-low attachment culture dish (Corning #CLS3261, Corning, NY, USA). Media were changed every day for 12 days till the neuroepithelial rosettes formed. The mature neuroectodermal spheres were then plated on a 1 μg/mL fibronectin (Sigma Aldrich #F1141) coated dish where the spheres attached, and migratory neural crest-like cells emerged.

The information on hiPSC and human islets was displayed in Supplementary Tables S1, S2. Human primary islets were maintained in CMRL 1066 (Corning, 15-110-CV) supplemented with 5% human AB serum (PAN-Niotech GmbH), L-Glutamine, 1% penicillin/streptomycin, 10 mM HEPES (all from Gibco) on ultra-low attachment plates (Corning, CLS3261). The hiPSCs were cultured in E8 Medium (Gibco, A1517001), and confirmed to be mycoplasma-free. The differentiation was done using the seven stages protocol [3 (link)] with modification [12 (link)]. On day 1 of the suspension culture, Rho Kinase inhibitor Y27632 (StemCell Technologies, 72304) was added to prevent cell death (Supplementary Table S3).