Identification and characterization of osteoclast-like cells and their progenitors in cultures of feline marrow mononuclear cells

K. J. Ibbotson, G. David Roodman, L. M. McManus, G. R. Mundy

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Abstract

The predominant cell reponsible for bone resorption, the multinucleated osteoclast, has been difficult to study because of inaccessibility. When feline marrow-derived mononuclear cells are established in long-term culture, multinucleated cells form within 48 h, reaching maximum numbers at 16 d. We have observed that these cultured cells have many of the features of osteoclasts. Morphologically, they are multinucleated, contain large numbers of branched mitochondria, have a peripheral cytoplasm lacking organelles (a clear zone), and have extensive cell-surface processes. In addition to these ultrastructural features, the cells contain a tartrate-resistant acid phosphatase, the activity of which is increased by parathyroid hormone (PTH) and inhibited by calcitonin. PTH, prostaglandin E2, and 1,25(OH)2 vitamin D3 increased multinucleated cell formation, while calcitonin inhibited the stimulatory effects of PTH. Time-lapse cinemicrographic and autoradiographic studies indicated that the multinucleated cells formed by fusion of the motonuclear progenitors. The multinucleated cells were phagocytic and stained with nonspecific esterase, consistent with their being derived from immature monocytes. Further, cell populations enriched for multinucleated cells release 45Ca from devitalized bone. Density-gradient centrifugation on Percoll was used to enrich and characterize the mononuclear progenitors of these multinucleated cells. The progenitor cells were found predominantly in Percoll density layers of 1.065 to 1.08 g/ml and were enriched up to 30-fold as compared to unfractionated cells. The bone marrow mononuclear cells that formed the multinucleated cells were initially nonadherent to plastic, stained heavily with nonspecific esterase, and appeared to be immature monocytes histologically. These data indicate that the multinucleated osteoclast-like cells in our cultures are devived from nonadherent monocytic progenitor cells that are responsive to osteotropic hormones. The ability to grow and characterize these cells in vitro should facilitate studies to elucidate the role these cells play in normal and pathologic states of bone resorption.

Original languageEnglish
Pages (from-to)471-480
Number of pages10
JournalJournal of Cell Biology
Volume99
Issue number2
StatePublished - 1984
Externally publishedYes

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Felidae
Osteoclasts
Stem Cells
Bone Marrow
Parathyroid Hormone
Carboxylesterase
Calcitonin
Bone Resorption
Monocytes
Density Gradient Centrifugation
Cell Fusion
Cholecalciferol
Phagocytes
Dinoprostone
Bone Marrow Cells
Organelles
Plastics
Cultured Cells
Mitochondria
Cytoplasm

ASJC Scopus subject areas

  • Cell Biology

Cite this

Identification and characterization of osteoclast-like cells and their progenitors in cultures of feline marrow mononuclear cells. / Ibbotson, K. J.; Roodman, G. David; McManus, L. M.; Mundy, G. R.

In: Journal of Cell Biology, Vol. 99, No. 2, 1984, p. 471-480.

Research output: Contribution to journalArticle

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