Differential Elasticity Affects Lineage Segregation of Embryonic Stem Cells

The question of what guides lineage segregation is central to development, where cellular differentiation leads to segregated cell populations destined for specialized functions. Here, using optical tweezers measurements of mouse embryonic stem cells, we reveal a mechanical mechanism based on differential elasticity in the second lineage segregation of the embryonic inner cell mass into epiblast (EPI) cells, which will develop into the fetus, and primitive endoderm (PrE), which will form extraembryonic structures such as the yolk sac. Remarkably, we find that these mechanical differences already occur during priming, not just after a cell has committed to differentiation. Specifically, we show that PrE-primed cells exhibit significantly higher elasticity than EPI-primed cells, characterized by lower power spectrum scaling exponents, higher Young's modulus, and lower loss tangent. Using a model of two cell types differing only in elasticity, we show that differential elasticity alone is sufficient to lead to segregation between cell types, suggesting that the mechanical attributes of the cells contribute to the segregation process. Importantly, we find that this process relies on cellular activity. Our findings present differential elasticity as a previously unknown mechanical contributor to lineage segregation during embryo morphogenesis.