naa a22 4500 510739261 CHVBK 20180411083022.0 cr unu---uuuuu 180411e20130801xx s 000 0 eng 10.1007/s12015-012-9412-5 doi (NATIONALLICENCE)springer-10.1007/s12015-012-9412-5 Generation of Mouse and Human Induced Pluripotent Stem Cells (iPSC) from Primary Somatic Cells [Elektronische Daten] [I. Lorenzo, A. Fleischer, D. Bachiller] Cellular reprogramming consists of the conversion of differentiated cells into pluripotent cells; the so-called induced Pluripotent Stem Cells. iPSC are amenable to in vitro manipulation and, in theory, direct production of any differentiated cell type. Furthermore, iPSC can be obtained from sick individuals and subsequently used for disease modeling, drug discovery and regenerative treatments. iPSC production was first achieved by transducing, with the use of retroviral vectors, four specific transcription factors: Oct4, Klf4, Sox2 and c-Myc (OKSM), into primary cells in culture Takahashi and Yamanaka, (Cell 126(4):663-676, 2006). Many alternative protocols have since been proposed: repeated transfections of expression plasmids containing the four pluripotency-associated genes Okita et al. (Science 322(5903):949-953, 2008), lentiviral delivery of the four factors Sommer et al. (Stem Cells 27(3):543-549, 2009), Sendai virus delivery Fusaki et al. (Proceedings of the Japan Academy. Series B, Physical and Biological Sciences 85(8):348-362, 2009), removal of the reprogramming vectors by ‘piggyBac' transposition Woltjen et al. (Nature 458(7239):766-770, 2009); Kaji et al. (Nature 458(7239):771-775, 2009), Cre-recombinase excisable viruses Soldner et al. (Cell 136(5):964-977, 2009), episomal vectors Yu et al. (Science 324(5928):797-801, 2009), cell-penetrating reprogramming proteins Zhou et al. (Stem Cells 4(5):381-384, 2009), mammalian artificial chromosomes Hiratsuka et al. (PLoS One 6(10):e25961, 2011) synthetically modified mRNAs Warren et al. (Scientific Reports 2:657, 2012), miRNA Anokye-Danso et al. (Cell Stem Cell 8(4):376-388, 2009); however, although some of these methods are commercially available, in general they still need to attain the reproducibility and reprogramming efficiency required for routine applications Mochiduki and Okita (Biotechnol Journal 7(6):789-797, 2012). Herein we explain, in four detailed protocols, the isolation of mouse and human somatic cells and their reprogramming into iPSC. All-encompassing instructions, not previously published in a single document, are provided for mouse and human iPSC colony isolation and derivation. Although mouse and human iPSC share similarities in the cellular reprogramming process and culture, both cell types need to be handled differently. Springer Science+Business Media New York, 2012 Induced pluripotent stem cells nationallicence Cellular reprogramming nationallicence Primary keratinocytes nationallicence Primary mouse embryonic fibroblasts nationallicence Retrovirus nationallicence Feeder cells nationallicence Cell derivation nationallicence Lorenzo I. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut Fleischer A. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut Bachiller D. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/4(2013-08-01), 435-450 1550-8943 9:4<435 2013 9 12015 https://doi.org/10.1007/s12015-012-9412-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12015-012-9412-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lorenzo I. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut NATIONALLICENCE 700 1- Fleischer A. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut NATIONALLICENCE 700 1- Bachiller D. Caubet-Cimera Foundation, Centre for Advanced Respiratory Medicine, Crta. Sóller Km12, 07110, Bunyola, Illes Balears, Mallorca, Spain aut NATIONALLICENCE 773 0- Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/4(2013-08-01), 435-450 1550-8943 9:4<435 2013 9 12015 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer