Skin fibroblasts were reprogrammed to iPSCs using non‐integrative episomal vectors expressing Oct4, Sox2, Klf4, c‐myc, p53shRNA, and Lin28, as previously described (Kuebler et al, 2017 (link)) to generate 2–4 independent iPSCs clones per individual. THD patient‐specific and control iPSCs were maintained in mTeSR™ medium (STEMCELL technologies) and passaged once a week with EDTA onto Matrigel‐coated (Cultek) plates (Life technologies). The absence of episomal expression and endogenous pluripotency markers was evaluated as previously reported (Kuebler et al, 2017 (link)). In vitro differentiation toward the endoderm, mesoderm, and neuroectoderm was performed essentially as described (Martí et al, 2013 (link)). The iPSCs generated and characterized in this study were registered and deposited with the Spanish Bank of Stem Cells. Information on iPSC lines is provided in Table 1.
Tristán‐Noguero A., Fernández‐Carasa I., Calatayud C., Bermejo‐Casadesús C., Pons‐Espinal M., Colini Baldeschi A., Campa L., Artigas F., Bortolozzi A., Domingo‐Jiménez R., Ibáñez S., Pineda M., Artuch R., Raya Á., García‐Cazorla À, & Consiglio A. (2023). iPSC‐based modeling of THD recapitulates disease phenotypes and reveals neuronal malformation. EMBO Molecular Medicine, 15(3), e15847.
Other organizations :
Hospital Sant Joan de Déu Barcelona, Institut d'Investigació Biomédica de Bellvitge, Universitat de Barcelona, Bellvitge University Hospital, Center of Regenerative Medicine in Barcelona, Duran i Reynals Hospital, Institut d'Investigacions Biomèdiques de Barcelona, Consejo Superior de Investigaciones Científicas, Consorci Institut D'Investigacions Biomediques August Pi I Sunyer, Centro de Investigación Biomédica en Red de Salud Mental, Instituto de Salud Carlos III, Instituto Murciano de Investigación Biosanitaria, Centro de Investigación Biomédica en Red, Centre for Biomedical Network Research on Rare Diseases, Hospital Universitario Virgen de la Arrixaca, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, Institució Catalana de Recerca i Estudis Avançats, University of Brescia
Skin fibroblasts were reprogrammed to iPSCs using non-integrative episomal vectors expressing Oct4, Sox2, Klf4, c-myc, p53shRNA, and Lin28
dependent variables
Generation of 2-4 independent iPSCs clones per individual
Absence of episomal expression and endogenous pluripotency markers
In vitro differentiation toward the endoderm, mesoderm, and neuroectoderm
control variables
THD patient-specific and control iPSCs were maintained in mTeSR™ medium (STEMCELL technologies) and passaged once a week with EDTA onto Matrigel-coated (Cultek) plates (Life technologies)
controls
Positive control: None specified
Negative control: None specified
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