title: Comparative analysis of targeted differentiation of human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells reveals variability associated with incomplete transgene silencing in retrovirally derived hiPSC lines.
abstract: Functional hepatocytes, cardiomyocytes, neurons, and retinal pigment epithelial (RPE) cells derived from human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) could provide a defined and renewable source of human cells relevant for cell replacement therapies, drug discovery, toxicology testing, and disease modeling. In this study, we investigated the differences between the differentiation potentials of three hESC lines, four retrovirally derived hiPSC lines, and one hiPSC line derived with the nonintegrating Sendai virus technology. Four independent protocols were used for hepatocyte, cardiomyocyte, neuronal, and RPE cell differentiation. Overall, cells differentiated from hESCs and hiPSCs showed functional similarities and similar expression of genes characteristic of specific cell types, and differences between individual cell lines were also detected. Reactivation of transgenic OCT4 was detected specifically during RPE differentiation in the retrovirally derived lines, which may have affected the outcome of differentiation with these hiPSCs. One of the hiPSC lines was inferior in all directions, and it failed to produce hepatocytes. Exogenous KLF4 was incompletely silenced in this cell line. No transgene expression was detected in the Sendai virus-derived hiPSC line. These findings highlight the problems related to transgene expression in retrovirally derived hiPSC lines.
authors: Research Programs Unit, Molecular Neurology, Biomedicum Stem Cell Center, University of Helsinki, Helsinki, Finland.
authors: Sanna
authors: S
authors: Toivonen
authors: Marisa
authors: M
authors: Ojala
authors: Anu
authors: A
authors: Hyysalo
authors: Tanja
authors: T
authors: Ilmarinen
authors: Kristiina
authors: K
authors: Rajala
authors: Mari
authors: M
authors: Pekkanen-Mattila
authors: Riikka
authors: R
authors: Äänismaa
authors: Karolina
authors: K
authors: Lundin
authors: Jaan
authors: J
authors: Palgi
authors: Jere
authors: J
authors: Weltner
authors: Ras
authors: R
authors: Trokovic
authors: Olli
authors: O
authors: Silvennoinen
authors: Heli
authors: H
authors: Skottman
authors: Susanna
authors: S
authors: Narkilahti
authors: Katriina
authors: K
authors: Aalto-Setälä
authors: Timo
authors: T
authors: Otonkoski
mesh_terms: D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D020868:Gene Silencing; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology; D009415:Nerve Net / Q000502:physiology; D009474:Neurons / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D055213:Retinal Pigment Epithelium / Q000166:cytology; D029082:Sendai virus / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics
chemical_list: C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human