title: Transgene-Free Disease-Specific iPSC Generation from Fibroblasts and Peripheral Blood Mononuclear Cells.
abstract: Induced pluripotent stem cells (iPSCs) offer great promise as tools for basic biomedical research, disease modeling, and drug screening. In this chapter, we describe the generation of patient-specific, transgene-free iPSCs from skin biopsies and peripheral blood mononuclear cells through electroporation of episomal vectors and growth under two different culture conditions. The resulting iPSC lines are characterized with respect to pluripotency marker expression through immunostaining, tested for transgene integration by PCR, and assayed for differentiation capacity via teratoma formation.
authors.affiliation: School of Medicine, Koç University, Istanbul, Turkey.
authors.forename: Kerem
authors.initials: K
authors.lastname: Fidan
authors.affiliation: School of Medicine, Koç University, Istanbul, Turkey.|Molecular Biology and Genetics Department, Faculty of Science, Istanbul University, Istanbul, Turkey.
authors.forename: Ayyub
authors.initials: A
authors.lastname: Ebrahimi
authors.affiliation: School of Medicine, Koç University, Istanbul, Turkey.
authors.forename: Özlem H
authors.initials: ÖH
authors.lastname: Çağlayan
authors.affiliation: School of Medicine, Koç University, Istanbul, Turkey.
authors.forename: Burcu
authors.initials: B
authors.lastname: Özçimen
authors.affiliation: School of Medicine, Koç University, Istanbul, Turkey. tonder@ku.edu.tr.
authors.forename: Tamer T
authors.initials: TT
authors.lastname: Önder
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D065150:Cellular Reprogramming; D003094:Collagen / Q000737:chemistry; D015925:Cryopreservation; D004338:Drug Combinations; D018274:Electroporation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007797:Laminin / Q000737:chemistry; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D011509:Proteoglycans / Q000737:chemistry; D011725:Pyridines / Q000494:pharmacology; D012867:Skin / Q000166:cytology / Q000378:metabolism; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D019076:Transgenes
chemical_list: D000577:Amides; D015415:Biomarkers; D004338:Drug Combinations; D004791:Enzyme Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D011509:Proteoglycans; D011725:Pyridines; C059485:matrigel; C108830:Y 27632; D003094:Collagen
keywords: Characterization of pluripotency; Disease modeling; Episomal vector; Induced pluripotent stem cells; Transgene-free; iPSC