Expression of a mutant retinoic acid receptor beta alters lineage differentiation in mouse embryonic stem cells.

We have introduced 1 to 2 copies of a deletion mutant (beta Delta C) of the human retinoic acid receptor beta into mouse embryonic stem (ES) cells. The beta Delta C-expressing cells were 10 to 100 times less sensitive to RA-induced differentiation in comparison with their parental cells. In the presence of 10(-7) M RA in monolayer culture, they showed no growth arrest or differentiation, but remained pluripotent. Embryoid bodies (EBs) derived from beta Delta C-expressing cells differentiated into cardiomyocytes rather than neurons after treatment with 10(-6) M RA, and became neurons upon exposure to 10(-5) or 10(-4) M RA. Remarkably, after 10 passages of continuous culture in the presence of 10(-7) MRA, they still were able to form chimeras after injection into blastocysts. These data suggest that appropriate levels of normal retinoid receptors are crucial for lineage-specific differentiation of mouse ES cells in vitro. The beta Delta C mutant protein may prove to be useful in promoting "stemness" of ES cells in culture.