Inhibition of TGFβ Signaling Promotes Ground State Pluripotency

Seyedeh-Nafiseh Hassani, Mehdi Totonchi, Ali Sharifi-Zarchi, Sepideh Mollamohammadi, Mohammad Pakzad, Sharif Moradi, Azam Samadian, Najmehsadat Masoudi, Shahab Mirshahvaladi, Ali Farrokhi, Boris Greber, Marcos J. Araúzo-Bravo, Davood Sabour, Mehdi Sadeghi, Ghasem Hosseini Salekdeh, Hamid Gourabi, Hans R. Schöler, Hossein Baharvand

Embryonic stem (ES) cells are considered to exist in a ground state if shielded from differentiation triggers. Here we show that FGF4 and TGFβ signaling pathway inhibitors, designated R2i, not only provide the ground state pluripotency in production and maintenance of naïve ES cells from blastocysts of different mouse strains, but also maintain ES cells with higher genomic integrity following long-term cultivation compared with the chemical inhibition of the FGF4 and GSK3 pathways, known as 2i. Global transcriptome analysis of the ES cells highlights augmented BMP4 signaling pathway. The crucial role of the BMP4 pathway in maintaining the R2i ground state pluripotency is demonstrated by BMP4 receptor suppression, resulting in differentiation and cell death. In conclusion, by inhibiting TGFβ and FGF signaling pathways, we introduce a novel defined approach to efficiently establish the ground state pluripotency.