Uterus provides an essential place for embryo/fetus to grow inside mother’s body. The study of tissue engineering approaches for uterine regeneration, however, is still limited compared to other organs. In this study, we developed a novel method to fabricate scaffold-free tissue with human endometrial stromal cells (hESCs). We utilized Y27632, ROCK inhibitor, to retain its shape in 3D structure. As well as immunohistochemical assay and Real time-PCR, we also examined its role for early implantation events using rat embryo. It will be expected to imply its potential usage as scaffold in uterine tissue engineering.