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Transgene-Free Disease-Specific iPSC Generation from Fibroblasts and Peripheral Blood Mononuclear Cells.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2016; Vol. 1353, pp. 215-31. - Publication Year :
- 2016
-
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.
- Subjects :
- Amides pharmacology
Animals
Biomarkers metabolism
Biopsy
Cell Differentiation drug effects
Collagen chemistry
Cryopreservation
Drug Combinations
Electroporation
Enzyme Inhibitors pharmacology
Fibroblasts drug effects
Fibroblasts metabolism
Gene Expression
Genetic Vectors chemistry
Genetic Vectors metabolism
Humans
Induced Pluripotent Stem Cells drug effects
Induced Pluripotent Stem Cells metabolism
Intercellular Signaling Peptides and Proteins pharmacology
Laminin chemistry
Leukocytes, Mononuclear drug effects
Leukocytes, Mononuclear metabolism
Mice
Plasmids genetics
Plasmids metabolism
Primary Cell Culture
Proteoglycans chemistry
Pyridines pharmacology
Skin cytology
Skin metabolism
Teratoma genetics
Teratoma metabolism
Teratoma pathology
Transgenes
Cell Culture Techniques methods
Cellular Reprogramming
Fibroblasts cytology
Induced Pluripotent Stem Cells cytology
Leukocytes, Mononuclear cytology
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1353
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 26126451
- Full Text :
- https://doi.org/10.1007/7651_2015_278