Mesenchymal Stem Cell (MSC) is a source of stem cells are multipotent that can be differentiated into many tipe of cells. MSC can be obtained from various sources one of which is from the bone marrow. But the number of MSCs in the bone marrow is very limited, thus requiring the production of MSC in vitro to obtain considerable amounts on the application or further research. The success of the production is very dependent on various factors such as isolation methods and the use of appropriate culture medium. This study aims to compare the Dulbecco's Modified Eagle's Medium (DMEM) and Minimum Essential Medium Eagle (MEM) as a culture medium producing MSC were isolated from bone marrow of mice tibia and femur using flushing methode. Research conducted at the Laboratory of stem cells, Center for Biomedical and Basic Technology of Health. Tibia and femur of mice cleared of fat and decontaminated using alcohol 70% for 2 minutes. Once clean both ends of the bone is cut and in flushing using 1 cc needle with DMEM culture medium and MEM. Medium replacement is done after 48 hours of culture. The results showed that the MSC yield from tibia and femur bone marrow is higher when cultured with MEM rather than DMEM (p <0.05). In this study the production of MSC from bone marrow of mice can be performed by using MEM medium with flushing method.