Generation of two heterozygous MYBPC3 mutation-carrying human iPSC lines, SCVIi001-A and SCVIi002-A, for modeling hypertrophic cardiomyopathy

Geometry and length control of 3D engineered heart tissues using direct laser writing

“…25 Results from a second cell line, GSB-L550 (SCVIi001-A) from Greenstone Biosciences, which is created from a patient with hypertrophic cardiomyopathy, are included for comparison in Fig. S8 † 26 . HiPSCs from the second cell line were maintained in complete mTeSR1 medium (Stem Cell) on Matrigel (Fisher) mixed 1 : 80 in DMEM/F-12 (Fisher) and split using Accutase (Sigma) at 70–90% confluence.…”

“…S8. † 26 HiPSCs from the second cell line were maintained in complete mTeSR1 medium (Stem Cell) on Matrigel (Fisher) mixed 1 : 80 in DMEM/F-12 (Fisher) and split using Accutase (Sigma) at 70–90% confluence. Once hiPSCs reached >90% confluence, they were differentiated to the cardiomyocyte lineage in RPMI 1640 medium (Gibco) supplemented with B27 minus insulin (ThermoFisher) and 1× GlutaMax (Fisher) by sequential targeting of the WNT pathway – activating the WNT pathway using 12 μM CHIR99021 (Tocris) for 24 hours for PGP1, and 6 μM CHIR99021 (Tocris) for 48 hours for GSB-L550 cell lines, and inhibiting the WNT pathway using 5 μM of IWP4 (Tocris) for 48 hours on Day 3 for PGP1 and Day 2 for GSB-L550 cell lines.…”

Generation of human induced pluripotent stem cell lines carrying heterozygous PLN mutation from dilated cardiomyopathy patients

“…Peripheral blood mononuclear cells (PBMCs) were isolated from patients’ blood by Percoll R gradient separation. PBMCs were purified and replated as previously described ( Liu et al, 2021 ). Briefly, PBMCs were cultured in 1 ml of Stem-Pro™-34 medium (100 ng/ml FLT3, 20 ng/ml IL-6, 20 ng/ml EPO, 20 ng/ml IL-3, and 100 ng/ml SCF).…”

“…We derived two human iPSC lines (SCVIi049-A and SCVIi050-A) from peripheral blood mononuclear cells (PBMCs) and fibroblasts of two patients carrying variants in the PLN gene, including a 44-year-old East Asian male (SCVIi049-A, c.25 C>T encoding p.Arg9Cys, likely pathogenic), and a 30-year-old Caucasian male (SCVIi050-A, c.40_42delAGA encoding p.Arg14del, pathogenic) ( Resource Table ). Reprogramming of somatic donor cells to iPSCs was conducted using a non-integrating Sendai virus containing the four Yamanaka factors described previously ( Liu et al, 2021 ). Both iPSC lines showed typical morphology ( Fig.…”

Generation of three heterozygous KCNH2 mutation-carrying human induced pluripotent stem cell lines for modeling LQT2 syndrome

“…PBMCs were isolated from blood by Percoll R separation, purified using DPBS and plated in a 24-well plate as previously described ( Liu et al, 2021 ). PBMCs (~1–2 × 10 6 ) were cultured in 1 mL of complete Stem-Pro ™ -34 medium combined with its specific supplements (in ng/mL): 100 FLT3, 20 IL-6 and 20 EPO, 20 IL-3 and 100 SCF as previously described ( Liu et al, 2021 ). Stem-Pro ™ -34 medium was changed every two days until cell culture stabilization.…”