Osteoclast formation in vitro from progenitor cells present in the adult mouse circulation

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Abstract
The development of multinucleated cells with tartrate-resistant acid phosphatase (TRAP) activity was studied in coverslip cultures of murine blood leukocytes and in cocultures of blood leukocytes with murine fetal bone rudiments. Cells with TRAP activity were not present among the leukocytes before culture and were absent in the bone rudiments at the time of explantation. After 14 days, macrophages with only tartrate-sensitive acid phosphatase activity developed in cultures of leukocytes without long bones. Multinucleated cells were not seen. In cocultures of leukocytes with bone rudiments, however, multinucleated cells with a strong TRAP activity had formed after 10–14 days of coculture. These TRAP-positive cells had invaded the bones and resorbed part of the calcified matrix. Electron microscopy revealed ruffled borders on the resorbing cells. In cocultures, TRAP-positive cells also formed from leukocyte fractions depleted of strongly adherent cells. Also on the cellophane supports of the cocultures, mononuclear cells with a stellate appearance and a strong TRAP activity were seen. We suggest that, in the cocultures, osteoclasts developed from a TRAP-negative, circulating progenitor cell. The presence of osteoclast progenitor cells in the circulation is discussed in light of the descent of osteoclasts from hematopoietic stem cells. That appearance of TRAP activity was always seen in resorbing cells and was not acquired in monocytes present in the leukocyte fraction by mere culture means that in the mouse TRAP is a useful marker for osteoclasts.
Funding Information
  • Foundation for Medical Research (900-541-069)