A system for culture of human trabecular bone and hormone response profiles of derived cells

Specimens of human trabecular bone from II patients were processed for tissue culture. In 10 out of 11 samples both cellular and matrix outgrowths were noted at the surfaces of explanted fragments after the first week in culture. During the second week adherent cells extended beyond the margins of the bone fragments and appeared to replicate. Plates achieved confluence in 30-36 days and cells were subcultured. In passaged cells doubling times were 5-7 days. Six cell cultures were examined for the presence of alkaline and acid phosphatase activity employing histochemical techniques. All six cultures contained cells which stained positively for alkaline phosphatase (10-80%). A small number of cells in one culture demonstrated tartrate-resistant acid phosphatase activity. Responses to hormones known to regulate the biological activities of skeletal tissues were also tested. Intracellular cyclic AMP was significantly increased by parathyroid hormone in three cultures, by salmon calcitonin in three cultures and by prostaglandin E2 in all 10 cultures. All three hormones increased the cyclic AMP content of cells cultured from human periosteum. It is concluded that cells cultured by this method demonstrate biochemical and morphological characteristics consistent with a skeletal tissue origin. Furthermore, such an approach may permit isolation and further characterization of individual subpopulations of bone cells of human origin.