Illumina HiSeq 2500 sequencing; 3'SRP data of cardiac differentiation of human induced pluripotent stem cell control lines
3'SRP data of cardiac differentiation of human induced pluripotent stem cell control lines
Directed cardiac differentiations of hiPSCs were performed using the established matrix sandwich method (Es-Salah-Lamoureux et al., 2016). Briefly, when hiPSCs reached 90% confluency, an overlay of Growth Factor Reduced Matrigel (0.033 mg/ml, BD Corning) was added. Differentiation was initiated 24h later by culturing the cells in RPMI1640 medium (Life Technologies) supplemented with B27 (without insulin, Life Technologies), 2 mM L-glutamine (Life Technologies), 1% NEAA (Life Technologies), 100 ng/mL Activin A (Miltenyi Biotec), 1X Pen/Strep (Life Technologies) and 10 ng/mL FGF2 for 24 hours. On the next day, the medium was replaced by RPMI1640 medium supplemented with B27 without insulin, 2 mM L-glutamine, 1% NEAA, 10 ng/mL BMP4 (Miltenyi Biotec), 1X Pen/Strep and 5 ng/mL FGF2 for 4 days. By day 5, cells were cultured in RPMI1640 medium supplemented with B27 complete (Life Technologies), 2 mM L-glutamine, 1X Pen/Strep and 1% NEAA and changed every two days until day 30. For each hiPSC line, samples were harvested daily from D-1 to D30 of the cardiac differentiation protocol from three independent cardiac differentiations. Total RNA were extracted using the NucleoSpin RNA kit (MACHEREY-NAGEL) and their quality assessed by NanoDropTM 1000 Spectrophotometer (Thermo Fisher Scientific). 3'RNA libraries were prepared by GenoBiRD core facility according to their published method (Charpentier et al., 2021)