@inbook{bad0afd238354ed9a997e39ae57626b2,
title = "Generation of Human iPSCs by Episomal Reprogramming of Skin Fibroblasts and Peripheral Blood Mononuclear Cells",
abstract = "Human-induced pluripotent stem cells (iPSCs) can be generated from patient-specific somatic cells by forced expression of the transcription factors OCT4, SOX2, KLF4, and c-MYC. Sustained expression of the transgenes during reprogramming is crucial for the successful derivation of iPSCs. Integrating retroviruses have been used to achieve the required prolonged expression; however, issues of undesirable transgene expression in the iPSC-derived cell types post reprogramming can occur. Alternative non-integrating approaches to reprogram somatic cells into pluripotency have been established. Here, we describe a detailed method for generating human iPSCs from fibroblasts and peripheral blood mononuclear cells (PBMCs) using the non-integrating episomal plasmids. The delivery of the episomal plasmids into the somatic cells is achieved using a nucleofection technique, and reprogramming is performed in chemically defined media. This process takes approximately 30 days to establish the iPSC colonies. We also describe a method for growing iPSCs on vitronectin as well as procedures for the long-term expansion of iPSCs on human fibroblast feeder cells.",
keywords = "Episomal reprogramming, Human peripheral mononuclear cells, Induced pluripotent stem cells, Stem cells, iPSC",
author = "Fabia Febbraro and Muwan Chen and Mark Denham",
year = "2021",
doi = "10.1007/978-1-0716-1084-8_9",
language = "English",
isbn = "978-1-0716-1083-1",
series = "Methods in Molecular Biology",
publisher = "Springer",
pages = "135--151",
editor = "Kejin Hu",
booktitle = "Nuclear Reprogramming",
address = "Germany",
}