Differentiating mouse ESC towards gonadal progenitors cells

We developed a gradual differentiation protocol to differentiate mESC towards the EpiSC lineage, followed by early mesoderm cells. These were later differentiated to intermediate mesoderm and we found a condition (IM3) that best generates early gonadal progenitor cells which are normally present at the bipotential gonad at E11.5. Forced expression of 2 transcription factors: Nr5A1 (SF1) and Dmrt1 induced a specific-Sertoli marker (TESCO-CFP) and the cells started to self-aggregate and form tubule-like structures, highly resembling the testis cords. There was proper induction of relevant markers at the correct time-points as shown by both qPCR, immuno-staining and RNA-Seq that includes comparison to in-vivo gonads. Overall design: Bulk RNA-Seq of cells at different stages of the differentiation: ESC, EpiSC, Early Mesoderm, Intermediate mesoderm, induced embryonic Sertoli like cells.