Reprogramming Methods Do Not Affect Gene Expression Profile of Human Induced Pluripotent Stem Cells

• Published in: International journal of molecular sciences 2017-01, Vol.18 (1), p.206-206
• Main Authors: Trevisan, Marta, Desole, Giovanna, Costanzi, Giulia, Lavezzo, Enrico, Palù, Giorgio, Barzon, Luisa
• Format: Article
• Language: English
• Subjects: Alkaline phosphatase; Animals; Cell Differentiation - genetics; Cell Line; Cells, Cultured; Cellular Reprogramming - genetics; Cellular Reprogramming Techniques - methods; Cloning; DNA microarrays; episomal vector; Fibroblasts - cytology; Fibroblasts - metabolism; Gene expression; Genetic Vectors - genetics; Humans; induced pluripotent stem cells; Induced Pluripotent Stem Cells - metabolism; Mice; Oligonucleotide Array Sequence Analysis; Plasmids - genetics; Pluripotency; Pluripotent Stem Cells - metabolism; reprogramming method; retroviral vector; Reverse Transcriptase Polymerase Chain Reaction; Sendai virus; Sendai virus - genetics; Sendai virus vector; Somatic cells; Stem cells; Transcriptome
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms18010206

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.