Fig. 1

Thyroid hormone T3 improves hPSC high-density survival and cloning efficiency. A Concentration of thyroid hormone T3 and T4 in different medium supplements at commonly used concentrations, measured by ELISA. n = 3 biological repeats. B, C Impact of T3 on cell apoptosis in high- and low-density culture treated with or without T3 for 3 days, measured with caspase 3/7 green detection reagent as shown in fluorescence images (B) and FACS data (C). Scale bar, 100 μm. Data are representative of three independent experiments. D T3 enhanced hESC survival after passaging from high-density culture. H1 cells were cultured in E8 or E8 + 500 nM T3 for 3 days until cells reach over 90% confluence, then passaged to a new plate and counted on the next day. n = 3 biological repeats, *P < 0.05. E, F T3 improved cloning efficiency. H1 cells were cultured in E8 or E8 + 500 nM T3 for 3 days and then collected using EDTA for seeding. 500 cells were seeded in each well of a 12-well plate and cultured in E8 or E8 with 500 nM T3 for 6 days. Alkaline phosphatase staining was used to visualize (E) and count the colonies (F). Scale bar, 5 mm. n = 3 biological repeats, *P < 0.05