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

Marta Trevisan, Giovanna Desole, Giulia Costanzi, Enrico Lavezzo, Giorgio Palù, and Luisa Barzon*
Source: Int J Mol Sci
Publication Date: (2017)
Issue: 18(1): 206
Research Area:
Stem Cells
Gene Expression
Cells used in publication:
Fibroblast, dermal (NHDF-Neo), human neonatal
Species: human
Tissue Origin: dermal
Fibroblast, dermal(NHDF-Ad), human adult
Species: human
Tissue Origin: dermal
Nucleofector® I/II/2b
4D-Nucleofector® X-Unit
In 2017,Trevisan et al published a very interesting publication, where the researchers compared retroviral vector systems, sendai virus based methods and the non-viral Nucleofection™ technology for reprogramming of adult somatic cells. The group analyzed 5 clones of each method with microarrays of 41,000 targets. The study showed that three different reprogramming strategies to obtain hiPSCs from differentiated cells show very similar expression profiles. That means that the method does not have an impact on the generated iPSCs. However, in their discussion section, the group examined many other aspects of the application methods. E.g. the efficiency for reprogramming fibroblast was most efficient with an episomal vector, and the viral vectors showed variances in reproducibility and safety concerns for generation of viruses and any future applications in humans.
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.