MSCs were initially obtained from bone marrow, but they can also be derived from other sources, such as skeletal muscle, umbilical cord blood, dental pulp, adipose tissue and amniotic fluid. MSCs have been successfully isolated and expanded from human, rat, rabbit, canine, pig and mouse. Mouse is the most widely used species in laboratory research because they are easy to manipulate and their genetic information is readily available. However, murine is the most difficult species to establish MSCs from BM. Murine BM is composed of heterogeneous cell populations that contain few MSCs. In addition, BMMSCs are located near the inner surface of the bone, making it difficult to flush them out. Another problem in establishing mouse BMMSCs is contamination with large amount of hematopoietic cells. Therefore, it is necessary to expand MSCs ex vivo. Such manipulation could cause cellular senescence by the loss of proliferation, differentiation and therapeutic potentials. This prompted us to look for an alternative source for MSCs with better ex vivo expansion capability. Endochondral ossification occurs during the process of long bone formation in foetal development. Primary ossification occurs at the bone centre for forming marrow cavity, while secondary ossification is formed in the bone epiphysis, followed by the formation of uncalcified cartilage, perichondrium and epiphyseal blood vessel penetration. Hence, we hypothesized the possibility of a biological niche for mesenchymal progenitors in the epiphysis. In this study, we derived novel MSCs from murine CUDC-907 epiphysis without enzymatic digestion. We characterized the morphology, proliferation and functional properties of EMSCs and compared these results with those of BMMSCs under the same cell culture conditions. We also evaluated the therapeutic effects of EMSCs on bone fracture and two types of ischemia mouse animal models. To our knowledge, this is a novel approach for the isolation of MSCs from murine bone. Although both EMSCs and BMMSCs were able to differentiate into SU5416 in vivo mesodermal cell lineages including adipocytes, osteocytes and chondrocytes, EMSCs showed higher potential to differentiate into these lineages. EMSCs showed greater potentials of proliferation and differentiation than BMMSCs, and also demonstrated paracrine antiinflammation ability by which derived various therapeutic effects of MSCs. To further confirm the potential of clinical application of EMSCs, we first investigated the osteogenesis potential of EMSCs using bone fracture model. EMSCs were seeded on a collagen-based gelatin sponge and implanted into the fracture site. 14 days after surgery, osteocalcification of the injured sites was evaluated by X-ray image, and significantly higher bone density was observed in the EMSCs treated group indicating that EMSCs can improve bone repair response.