Generation and Characterization of Human Pluripotent Stem Cells

H1 hESCs were purchased from WiCell Institute. The H1 SOX2-P2A-H2B-tdTomato reporter line was generated by knocking in a P2A-H2B-tdTomato transgene before the stop codon at the SOX2 gene locus using CRISPR/Cas9-based HDR³⁸ . The MSK-SRF001-iPSCs were generated from urine cells of an apparently healthy donor using a previously reported method^{39 (link)}. The 972-iPSCs were previously generated from an HGPS patient’s fibroblast line purchased from Coriell Institute (AG01972)^{40 (link)}, and the 756-iPSCs were generated from a PD patient’s fibroblast line purchased from Coriell Institute (ND29756). All the hPSCs have been fully characterized and are routinely cultured on Matrigel (Fisher Scientific 08-774-552) in Stemflex Medium (Thermo Fisher A3349401). Cells were maintained at 37 °C with 5% CO₂. For regular passaging, cells were detached and passaged with 0.5 mM EDTA (Fisher Scientific MT-46034CI) at room temperature for 5 min.