title: Reprogramming Methods Do Not Affect Gene Expression Profile of Human Induced Pluripotent Stem Cells.
abstract: Induced pluripotent stem cells (iPSCs) are pluripotent cells derived from adult somatic cells. After the pioneering work by Yamanaka, who first generated iPSCs by retroviral transduction of four reprogramming factors, several alternative methods to obtain iPSCs have been developed in order to increase the yield and safety of the process. However, the question remains open on whether the different reprogramming methods can influence the pluripotency features of the derived lines. In this study, three different strategies, based on retroviral vectors, episomal vectors, and Sendai virus vectors, were applied to derive iPSCs from human fibroblasts. The reprogramming efficiency of the methods based on episomal and Sendai virus vectors was higher than that of the retroviral vector-based approach. All human iPSC clones derived with the different methods showed the typical features of pluripotent stem cells, including the expression of alkaline phosphatase and stemness maker genes, and could give rise to the three germ layer derivatives upon embryoid bodies assay. Microarray analysis confirmed the presence of typical stem cell gene expression profiles in all iPSC clones and did not identify any significant difference among reprogramming methods. In conclusion, the use of different reprogramming methods is equivalent and does not affect gene expression profile of the derived human iPSCs.
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. marta.trevisan@unipd.it.
authors.forename: Marta
authors.initials: M
authors.lastname: Trevisan
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. giovanna.desole@studenti.unipd.it.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Desole
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. giulia.costanzi.gc@gmail.com.
authors.forename: Giulia
authors.initials: G
authors.lastname: Costanzi
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. enrico.lavezzo@unipd.it.
authors.forename: Enrico
authors.initials: E
authors.lastname: Lavezzo
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. giorgio.palu@unipd.it.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palù
authors.affiliation: Department of Molecular Medicine, University of Padova, 35121 Padova, Italy. luisa.barzon@unipd.it.
authors.forename: Luisa
authors.initials: L
authors.lastname: Barzon
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics*; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029082:Sendai virus / Q000235:genetics; D059467:Transcriptome
keywords: Sendai virus vector; episomal vector; gene expression; induced pluripotent stem cells; reprogramming method; retroviral vector