Thiazovivin

Human embryonic kidney 293T (HEK293T) cells were cultured in Dulbecco’s Eagle Medium (DMEM) (Life Technologies), 10% fetal bovine serum (FBS) (Gibco), and 1× Penicillin/Streptomycin (Life Technologies). HEK293T cells were from ATCC (CRL-3216).
Undifferentiated hPSCs were cultured in hPSC medium: DMEM/F12 (Life Technologies), 20% KnockOut Serum Replacement (KSR) (Life Technologies), 1× Non-Essential Amino Acids (NEAA) (Life Technologies), 0.055 mM 2-mercaptoethanol (Sigma), 1× Penicillin/Streptomycin, and 10 ng ml⁻¹ bFGF (Peprotech). hPSCs were maintained on CF1 feeder cells at 37 °C and 5% CO₂. hPSCs were isolated by Accutase (Life Technologies) as 1:3 to 1:6 every 3–6 days. 0.5 μM thiazovivin (TargetMol) were used during the first 24 h when passaging or thawing cells. MEL1 INS^GFP/W hESC line^{46 (link)} was a kind gift from Drs. E. G. Stanley and Andrew Elefanty. Mycoplasma contamination was routinely detected using TransDetect PCR Mycoplasma Detection Kit (TransGen Biotech).

Human embryonic kidney (HEK) 293T cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (FBS) (Gibco) and 1× penicillin/streptomycin (Life Technologies). HEK293T cells were bought from the American Type Culture Collection (ATCC) (CRL-3216).
Undifferentiated hESCs and hiPSCs were cultured in hPSC medium: DMEM/F12 (Life Technologies) supplemented with 20% KnockOut serum replacement (Life Technologies), 1× nonessential amino acids (Life Technologies), 1× penicillin/streptomycin, 0.055 mM 2-mercaptoethanol (Sigma-Aldrich), and bFGF (10 ng ml⁻¹; Peprotech). hPSCs were maintained on CF1 feeder cells in a humidified incubator at 37°C and 5% CO₂. hPSCs were passaged by Accutase (Life Technologies) as 1:3 to 1:6 every 3 to 6 days. Thiazovivin (0.5 μM; TargetMol) was used during the first 24 hours when passaging or thawing cells. MEL1 INS^GFP/W hESC line was a gift from E. G. Stanley and A. Elefanty. H9 hESCs were purchased from Wicell Research Institute. hiPSCs were produced by reprogramming human fibroblasts that were purchased from ATCC (CRL-2097) (49 (link), 52 (link)). Mycoplasma contamination was routinely detected using the TransDetect PCR Mycoplasma Detection Kit (TransGen Biotech). Institutional Animal Care and Use Committee guidelines were followed with human or animal subjects.