A morphologic study of osteoclasts isolated from osteopetrotic microphthalmic (mi/mi) mouse and human fetal long bones using an instrument permitting combination of light and scanning electron microscopy

Abstract

Cell surface structures of isolated osteopetrotic (mi/mi) and normal murine (+/+ and +/mi) and human osteoclasts were examined in a microscope combining light and scanning electron microscopy (LM/SEM). Tartrate-resistant acid phosphatase (TrAP) was used as an histochemical osteoclast marker. In osteopetrotic bone, as in normal murine bone, TrAP activity was exclusively seen in osteoclasts and preosteoclasts and was therefore judged a suitable marker for identification of isolated osteoclasts. A method was developed for preparation of LM/SEM specimens from osteoclast-enriched cell suspensions. In the LM/SEM isolated osteoclasts were easily recognized in the LM mode by TrAP contents. In specimens prepared from murine cells, but not human cells, LM identification of osteoclasts by TrAP was essential. This was in particular true for small, mononuclear, mi/mi osteoclasts. All osteoclasts examined had a villous appearance and were well spread over the glass substrate. There were no differences in cell surface morphology and in adherence to glass between osteopetrotic and normal osteoclasts.