Osseous tissue can be formed by culturing marrow cells with compounds such as dexamethasone and that a bone matrix cultured in this manner possesses BMP activity. We have reported that artificial bones with a high level of osteogenic potential can be prepared by culturing artificial bone materials with cultured osseous tissue. Here, in an attempt to develop activated cultured bone constructs with even greater osteogenic potential, the effects of the female hormone estriol on osteogenesis were investigated. Bone marrow cells were collected from the femur shafts of 7-week-old male Fischer rats, and subjected to primary and secondary cultures. During secondary culture with or without dexamethasone (Dx), 10-5, 10-6, 10-7, 10-8 or 10-9 M of estriol was added to a standard culture medium containing ascorbic acid and β-glycerophosphosphate. The alkaline phosphatase(ALP) activity and Ca levels were measured and statistically analyzed. There was a significant difference in ALP activity between the control group and the estriol groups, and ALP activity was the highest in the 10-7 and 10-8 M groups, being about 2.5 times higher than in the control group. Similar results were seen for Ca levels. Furthermore, in vivo study showed10-7M-estriol-treated-cultured bone/ceramic construct has significant high osteogenic potential when it is grafted into in vivo. Estriol has been reported to increase bone mass, and the results of the present study suggest that the osteogenic potential of cultured bone constructs can be more than doubled by adjusting the concentration of estriol in bone marrow cell culture. Therefore, the use of estriol may be able to facilitate osteogenesis in bone regeneration therapy.