Cytotune sendai virus

The 139 iPSCORE subjects [86 (link)] included members of 27 families (2–9 members/family), 7 monozygotic twin pairs, and 55 singletons (S1 Table). During recruitment of iPSCORE subjects, subject information (sex, age, family, ethnicity, and cardiac diseases) was collected. Recruitment was approved by the Institutional Review Boards of the University of California, San Diego and The Salk Institute (project no. 110776ZF). Skin biopsies were collected for each iPSCORE individual.
As previously described in detail [86 (link)–88 (link)], we reprogrammed skin fibroblast samples from 139 iPSCORE individuals using non-integrative Cytotune Sendai virus (Life Technologies) and the 149 iPSCs (7 subjects had 2 or more clones each) were shown to be pluripotent and to have high genomic integrity with no or low numbers of somatic copy-number variants (CNVs). To generate iPSC-derived cardiovascular progenitors (iPSC-CVPC) we used a small molecule differentiation protocol, which included purification of iPSC-CVPC cultures using lactate [89 (link)], and harvested iPSC-CVPC at day 25 (D25) differentiation [14 (link),85 (link)]. We obtained RNA-seq (150 bp PE) for 180 iPSC-CVPC (30 of the 149 iPSCs were differentiated two or more times). All iPSCORE iPSC-CVPC RNA-seq samples are available through dbGaP (phs000924).

As previously described, we reprogrammed fibroblast samples using non-integrative Cytotune Sendai virus (Life Technologies), and the 191 iPSCs (seven subjects had two or more clones each) were shown to be pluripotent and to have high genomic integrity with no or low numbers of somatic copy-number variants (CNVs) (D'Antonio et al., 2018 (link), Panopoulos et al., 2017a (link)).

Generation of the 106 iPSC lines has previously been described in detail^{39 (link)}. Briefly, cultures of primary dermal fibroblast cells were generated from a punch biopsy tissue^{100 (link)}, infected with the Cytotune Sendai virus (Life Technologies) per manufacturer’s protocol to initiate reprogramming. Emerging iPSC colonies were manually picked after Day 21 and maintained on Matrigel (BD Corning) with mTeSR1 medium (Stem Cell Technologies). Multiple independently established iPSC clones (i.e. referred to as lines) were derived from each individual. Many of the iPSC lines were evaluated by flow cytometry for expression of two pluripotent markers: Tra-1-81 (Alexa Fluor 488 anti-human, Biolegend) and SSEA-4 (PE anti-human, Biolegend)^{39 (link)}. Pluripotency was also examined using PluriTest-RNAseq^{39 (link)}. This iPSCORE resource was established as part of the Next Generation Consortium of the National Heart, Lung and Blood Institute and is available to researchers through the biorepository at WiCell Research Institute (www.wicell.org; NHLBI Next Gen Collection). For-profit organizations can contact the corresponding author directly to discuss line availability.