Human osteoblasts derived from long bone periosteum were induced to mineralize in culture in the presence of 2 mM alpha-glycerophosphate, with typical characteristics of mineralization, namely, accumulation of hydroxyapatite and increases in alkaline phosphatase activity and in osteocalcin production. Mineralization was also enhanced by 10(-8) M 1 alpha, 25-dihydroxyvitamin D3. In this system, a prostaglandin A1 derivative, TEI-3313, with the chemical structure 5-[(Z,2E)-4,7-dihydroxy-2-heptenyridene]-4-hydroxy-2-methylthio-4- (4- phenoxybutyl)-2-cyclopentenone, was found to enhance mineralization as effectively as 1 alpha, 25-dihydroxyvitamin D3, although its potency was 10 times lower than that of the vitamin D3 metabolite. Osteocalcin, a bone-specific noncollagenous matrix protein, accumulated onto the cell layers by treatment with TEI-3313 to a much greater extent than those released into the culture medium. TEI-3313 also enhanced collagen synthesis. Based on the finding that TEI-3313 enhanced the synthesis of both collagen and noncollagenous protein, it is speculated that TEI-3313 enhanced the mineralization by stimulating the expression of various genes in osteoblasts.