title: Practical Integration-Free Episomal Methods for Generating Human Induced Pluripotent Stem Cells.
abstract: The advent of induced pluripotent stem (iPS) cell technology has revolutionized biomedicine and basic research by yielding cells with embryonic stem (ES) cell-like properties. The use of iPS-derived cells for cell-based therapies and modeling of human disease holds great potential. While the initial description of iPS cells involved overexpression of four transcription factors via viral vectors that integrated within genomic DNA, advances in recent years by our group and others have led to safer and higher quality iPS cells with greater efficiency. Here, we describe commonly practiced methods for non-integrating induced pluripotent stem cell generation using nucleofection of episomal reprogramming plasmids. These methods are adapted from recent studies that demonstrate increased hiPS cell reprogramming efficacy with the application of three powerful episomal hiPS cell reprogramming factor vectors and the inclusion of an accessory vector expressing EBNA1.
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.|Present address: Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Cody
authors.initials: C
authors.lastname: Kime
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.
authors.forename: Tim A
authors.initials: TA
authors.lastname: Rand
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.|Department of Pediatrics, University of California, San Francisco, California.
authors.forename: Kathryn N
authors.initials: KN
authors.lastname: Ivey
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.|Department of Pediatrics, University of California, San Francisco, California.|Department of Biochemistry and Biophysics, University of California, San Francisco, California.
authors.forename: Deepak
authors.initials: D
authors.lastname: Srivastava
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.|Department of Anatomy, University of California, San Francisco, California.|Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.affiliation: Gladstone Institute of Cardiovascular Disease, San Francisco, California.|Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, California.
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Tomoda
mesh_terms: D018929:Cell Culture Techniques; D002469:Cell Separation; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics*; D014162:Transfection
keywords: episomal; human; induced pluripotent stem cell; reprogramming