268 Chapter 10 Table 2. Reagent Details Classification Result Data Morphology Brightfield imaging Fig. 1A Phenotype Immunocytochemistry RT-qPCR Positive for pluripotency markers NANOG and OCT4. Expression of pluripotency genes was assessed by qPCR Fig. 1D Fig. 1E Genotype Karyotype (ddPCR) Banding quality Supplementary Fig. 1 Identity Microsatellite PCR STR analysis Not performed 16 loci analyzed, ruled out crosscontamination N/A Not yet performed Mutation analysis TaqMan Genotype Assay Not performed N/A Microbiology and virology Mycoplasma MycoAlertTM Mycoplasma Detection Kit: All negative Data not shown Differentiation potential Directed differentiation All hiPSC lines showed full differentiation capacity towards mesodermal(cardiac) germ layer and CM expansion potential. Fig. 1B, 1C List of recommended germ layer markers Trilineage differentiation qPCR array Not performed N/A Donor screening (OPTIONAL) HIV 1 + 2 Hepatitis B, Hepatitis C Not performed N/A 4. Resource details PLN is a major regulator of calcium homeostasis in the SR and acts through the inhibition of the sarcoplasmic reticulum Ca2+ ATPase (SERCA).1 During β-adrenergic signalling, PLN is phosphorylated at Ser16 by PKA, or at Thr17 by calcium/calmodulin-dependent protein kinase II (CaMKII) or protein kinase B (Akt), which reverses the inhibitory effect on SERCA.2 Although the specific cardiac pathology and onset of disease vary, most patients present in the clinics with cardiac dilatation, hypertrophy, decreased ejection fraction, and ventricular arrhythmias. In this regard, the pathogenic c.40_42delAGA (p.Arg14del) mutation is associated with severe heart failure and a poor prognosis from late adolescence.3 Therefore, it is crucial to generate hiPSCs of patients with the PLN-R14del genetic mutation to elucidate the disease mechanism of this cardiomyopathy. This study establishes novel hiPSC, lines, of which six carry the genetic variant PLN-R14del c.40_42delAGA, where 3 carriers experienced cardiomyopathy-related symptoms.
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