Human osteoclast‐like cells in primary culture

Abstract

We describe a method for the isolation and culture of osteoclast-like cells from cancellous bone chips of iliac crests from patients undergoing reconstructive maxillofacial surgery. Under aseptic conditions, bone chips were cut into small pieces, incubated briefly with collagenase, and the isolated bone cells were separated from the bone chips by filtration using a nylon mesh. Bone cells were then cultured on a variety of surfaces for up to 10 days. Cell motility and fusion, together with the development of tartrate-resistant acid phosphatase activity, were seen in many cells soon after culture. The large osteoclast-like cells adhered to human cortical bone slices and produced resorption pits. These morphological and functional characteristics suggest that the cells we isolated and cultured were human osteoclasts and their precursors. Thus this method may provide a reliable means of obtaining human osteoclasts from normal tissue for short-term studies of their metabolism or from various skeletal diseases to study pathological aberrations and mechanisms. © 1996 Wiley-Liss, Inc.