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.
Keywords:
Characterization of pluripotency; Disease modeling; Episomal vector; Induced pluripotent stem cells; Transgene-free; iPSC.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amides / pharmacology
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Animals
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Biomarkers / metabolism
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Biopsy
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Cell Culture Techniques / methods*
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Cell Differentiation / drug effects
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Cellular Reprogramming*
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Collagen / chemistry
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Cryopreservation
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Drug Combinations
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Electroporation
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Enzyme Inhibitors / pharmacology
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Fibroblasts / cytology*
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Fibroblasts / drug effects
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Fibroblasts / metabolism
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Gene Expression
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Humans
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Induced Pluripotent Stem Cells / cytology*
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Induced Pluripotent Stem Cells / drug effects
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Induced Pluripotent Stem Cells / metabolism
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Intercellular Signaling Peptides and Proteins / pharmacology
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Laminin / chemistry
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Leukocytes, Mononuclear / cytology*
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Leukocytes, Mononuclear / drug effects
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Leukocytes, Mononuclear / metabolism
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Mice
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Plasmids / genetics
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Plasmids / metabolism
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Primary Cell Culture
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Proteoglycans / chemistry
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Pyridines / pharmacology
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Skin / cytology
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Skin / metabolism
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Teratoma / genetics
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Teratoma / metabolism
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Teratoma / pathology
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Transgenes
Substances
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Amides
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Biomarkers
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Drug Combinations
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Enzyme Inhibitors
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Intercellular Signaling Peptides and Proteins
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Laminin
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Proteoglycans
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Pyridines
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matrigel
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Y 27632
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Collagen