Abstract
The study of blastocyst differentiation from stem cells is still at its stage of infancy. In this study, a simple and efficient method was developed to derive blastocyst-like cell aggregates (BLCAs) from murine embryonic stem cells (mESCs). This was achieved by culturing the mESCs in non-adherent dishes.0020BLCAs (13±2 per million mESCs) were obtained in only 5 days and they were found to express the protein and gene markers of trophoblasts. Furthermore, the BLCAs showed the normal trophoblasts' capability of penetrating the uterine wall after being transferred into pseudo-pregnant mice, indicating the trophoblast-like functionality of the derived BLCAs. Collectively, we successfully developed a simple and fast 3D suspension culture approach to differentiate mESCs into BLCAs, which may provide important clues into the development of early embryos and stem cells. This may also offer a simple and efficient approach to isolate trophoblast cells for studying the native microenvironment of pluripotent stem cells without the need to sacrifice animals.