Abstract
Adult fibroblasts can be reprogrammed into induced pluripotent stem cells (iPSC) for use in various applications. However, there are challenges in iPSC generation including low reprogramming efficiency, yield, cell survival and viability. Since the Hippo signalling pathway is a key pathway involved in regulating cell proliferation and survival, we here test whether modification of the Hippo pathway will enhance the efficiency of iPSC generation and improve their survival.The Hippo pathway was modified by genetic ablation of the mammalian sterile-20 like kinase 1 (Mst1), a major component of the pathway. Using adult skin fibroblasts isolated from Mst1 knockout mice (Mst1−/−) as a source of iPSC we found that genetic ablation of Mst1 leads to significantly increased reprogramming efficiency by 43.8%. Moreover, Mst1−/− iPSC displayed increase proliferation by 12% as well as an increase in cell viability by 20% when treated with a chemical hypoxic inducer. Mechanistically, we found higher activity of YAP, the main downstream effector of the Hippo pathway, in iPSC lacking Mst1.In conclusion, our data suggests that Mst1 can be targeted to improve the efficiency of adult somatic cell reprogramming as well as to enhance iPSC proliferation and survival.
Highlights
Induced pluripotent stem cells have been used in an increasing number of applications since Takahashi and Yamanaka (Takahashi and Yamanaka, 2006) first demonstrated the reprogramming of adult somatic cells to produce iPSC
Adult skin fibroblasts were cultured from the ear skin tissue biopsies of 12 week old mammalian sterile-20 like kinase 1 (Mst1)−/− and wild type (WT) mice
We used an EdU incorporation assay to examine the effects of Mst1 ablation on cell proliferation rate
Summary
Induced pluripotent stem cells (iPSC) have been used in an increasing number of applications since Takahashi and Yamanaka (Takahashi and Yamanaka, 2006) first demonstrated the reprogramming of adult somatic cells to produce iPSC. In combination with gene editing technology iPSC have been widely used to model human genetic diseases (Avior et al, 2016), providing a useful alternative to animal models. IPSC-derived cells are valuable tools in drug discovery since these cells, which can carry phenotypes of a particular disease, can be used in high-throughput screening to find pharmacological compounds that may correct the phenotypes (Avior et al, 2016). A. Robertson et al / Stem Cell Research 20 (2017) 42–49 identification of novel reprogramming enhancers that are pharmacologically targetable would be very useful
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
