Hescs h1 and h9

To compare the metabolic differences between PSCs and somatic cells, hESCs (H1 and H9; WiCell Research Institute), hiPSCs (CMC-hiPSC-003 and CMC-hiPSC-009; (National Center for Stem Cell and Regenerative Medicine) and fibroblast cell lines (BJ1 and MRC5) were used. This study was approved by the Institutional Review Board (IRB No: P01-202309-03-001). These PSCs were maintained in complete TeSR-E8 Medium (STEMCELL Technologies) as the glucose-free culture. Cells were cultured on Vitronectin XF-coated plates (STEMCELL Technologies) and dissociated by incubation with TrypLE Express (Gibco) at 37 for 3 minutes. The fibroblast cell lines, BJ1 and MRC5, were maintained in Dulbecco's modified Eagle medium (high-glucose DMEM; Corning Life Sciences) supplemented with 10% fetal bovine serum (FBS; Corning Life Sciences) and 100× penicillin-streptomycin (Corning Life Sciences). hESCs and hiPSCs were routinely passaged every 3 days using TrypLE Express, whereas fibroblasts were passaged every 3 5 days using 0.05% trypsin-ethylenediaminetetraacetic acid. All cells were maintained in an incubator at 37 and 5% CO 2 .

Human ESCs (hESCs), H1 and H9, were obtained from WiCell Research Institute (Maddison, USA) and HUES8 was obtained from Harvard University. hESCs were cultured and differentiated into pancreatic progenitors as previously reported. 31, 32 For differentiation into beta cells, we used two different protocols with slight modifications 24, 25 (Supplementary Materials; Table S1).
Immunostaining and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were performed as previously described. 31, 32 The antibodies and primers used in this study are listed in Tables S2 andS3.