Epigenetic Modifiers Facilitate Induction and Pluripotency of Porcine iPSCs

Inadequate silence of exogenous genes represents a major obstacle to complete epigenetic reprogramming of pig induced pluripotent stem cells (piPSCs) by conventional pluripotency trasncription factors. We tested the hypothesis that epigenetic modification by active DNA or histone demethylation or by inhibition of histone deacetylase would enhance reprogramming and exogenous gene silencing in piPSCs. piPSCs induced by OSKM in combination with epigenetic factors, specifically Ten-Eleven-Translocation (Tet1 or Tet3) and lysine (K)-specific demethylase 3A (Kdm3a), expressed higher levels of Rex1 and other genes representing naïve state and exhibited more open chromatin status, in contrast to those of OSKM controls. Moreover, piPSCs induced by Tet1 combined with OSKM exhibited enhanced differentiation capacity. Conversion with histone deacetylase inhibitors, including NaB, TSA and VPA further increased expression of Rex1 and reduced expression of exogenous genes, generating high pluripotent piPSCs. Together, epigenetic modifiers can enhance generation of piPSCs and reduce their reliance on exogenous genes. RNA-seq for progenitor PEF, piPSCs P5 (Rex1+) and piPSCs P10 (Rex1-), in duplicate.