br Materials and methods br Acknowledgements We

Materials and methods
Acknowledgements We would like to thank Dr. Keisuke Okita and Prof. Shinya Yamanaka for providing the plasmids. Furthermore, we would like to thank Prof. Duncan R. Smith for proof reading the article. This work was supported by the Thailand Research Fund (MRG5980187), National Research Council of Thailand (NRCT; DPAR/2559-158), the Office of the Higher Education Commission and Mahidol University under the National Research University Initiative, a Research Chair Grant from the National Science and Technology Development Agency (NSTDA; P-11-00435), and European Union 7th Framework Program (PIAPP-GA-2012-324451-STEMMAD). Wasinee Wongkummool is supported by the Thailand Graduate Institute of Science and Technology (TGIST; TG-22-14-5-036D) scholarship, NSTDA.
Resource table Resource details Peripheral blood CD34+ hematopoietic progenitor (HP) cells were collected from a 52year old female patient who has been diagnosed with homozygous HbCS. The peripheral blood CD34+ HPs were then reprogrammed into a human induced pluripotent stem cell line by co-electroporation of episomal plasmids expressing hOCT4, hSOX2, hL-MYC, hKLF4, hNANOG, hLIN28, a short hairpin RNA against TP53 and Epstein-Barr nuclear antigen-1 (EBNA-1) (Okita et al., 2011). The iPSC like colonies were picked 21–28days post electroporation (Fig. 1A). The absence of the reprograming plasmids in the genome was verified by PCR after 10 passages in the MUi017-A cell line (Fig. 1B and Supplementary Fig. 1). Multiplex GAP-polymerase chain reaction (Gap-PCR) and sequencing analysis confirmed the absence of four common α-globin gene deletions [–α (rightward), –α4.2 (leftward) --SEA and --THAI] and a homozygous c.427 (T>C) StemRegenin 1 Supplier substitution in termination codon of HBA2 gene respectively (Fig. 1C and D). The cells expressed the pluripotency markers NANOG, OCT4, TDGF1, DNMT3B, GABRB3, SOX2, and GDF3 in the same range as MU011.A-hiPSC, a control iPSC line described previously (Tangprasittipap et al., 2015) (Fig. 1E). Expression of the pluripotency markers, OCT4 and NANOG at the protein level was confirmed by immunofluorescence staining (Fig. 1F). In vitro differentiation followed by immunofluorescent staining analysis with the ectodermal marker beta-III-Tubulin (TUJ1), the mesodermal marker smooth muscle actin (SMA) and the endodermal marker alpha-fetoprotein (AFP) demonstrated the potential for differentiation into cells derived from all three germ layers (Fig. 1G) In addition, the iPSCs presented a normal female karyotype (46, XX) (Fig. 1H). DNA fingerprinting of MUi017-A cell line is identical to their parental cells (Supplementary Table 1) and mycoplasma infection was negative by PCR (Supplementary Fig. 2).
Materials and methods
Acknowledgements We would like to thank Dr. Keisuke Okita and Prof. Shinya Yamanaka for providing the plasmids. This work was supported by the Thailand Research Fund (MRG5980187), National Research Council of Thailand (NRCT; DPAR/2559-158), the Office of the Higher Education Commission and Mahidol University under the National Research University Initiative, a Research Chair Grant from the National Science and Technology Development Agency (NSTDA; P-11-00435), and European Union Seventh Framework Programme (PIAPP-GA-2012-324451-STEMMAD). Wasinee Wongkummool is supported by the Thailand Graduate Institute of Science and Technology (TGIST; TG-22-14-57-036D) scholarship, NSTDA.
Resource table Resource details Hypertrophic cardiomyopathy (HCM) is one of the most common genetic heart diseases with a prevalence of up to 1 in 200 people (Semsarian et al., 2015). Clinically, HCM is characterised by unexplained hypertrophy of the left ventricle (maximum left ventricular wall thickness≥15mm). Variants in myosin-binding protein C (MYBPC3) and beta-myosin heavy chain are the most common genetic cause of HCM; together they account for approximately 70% of all pathogenic variants (Maron et al., 2014). The pathogenesis of HCM remains largely unknown. However, studies combining induced pluripotent stem cell (iPSC) technology with genetic test results are starting to uncover mechanisms that may be involved in the development of HCM and other inherited cardiomyopathies (Ross et al., 2016).