In vitro differentiation protocol of Paraxial mesoderm from mouse embryonic stem cells (mESCs)

Stem cell-derived tissues have wide potential for modelling developmental and pathological processes as well as cell-based therapy. However, it has proven difficult to generate several key cell types in vitro, including skeletal muscle. In vertebrates, skeletal muscles derive during embryogenesis from the presomitic mesoderm (PSM). Using PSM development as a guide, we establish conditions for the differentiation of monolayer cultures of mouse embryonic stem (ES) cells into PSM-like cells without the introduction of transgenes or cell sorting. We differentiated mouse ESCs in a medium supplemented with Rspo3, DMSO and the Bmp inhibitor Noggin. In vivo, the PSM cells are first expressing Msgn1 (posterior PSM marker) . After 3 and 4 days of differentiation of mESCs, Msgn1-positive cells were FACS-sorted and their transcriptome analyzed.

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