Ultrastructural characterization of preosteoclasts derived from bone marrow progenitors stimulated by osteoclast colony stimulating factor

Lee, Minako Y.; Jonas, Mechthild; Lottsfeldt, Joan L.; Emil, Y.

doi:10.1002/(sici)1097-0185(199610)246:2<176::aid-ar4>3.0.co;2-#

Cited by 2 publications

(1 citation statement)

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“…These colonies were composed of mononuclear cells strongly positive for tartrate‐resistant acid phosphatase (TRAPase), a cytochemical enzyme marker for the osteoclast. Osteoclastic properties of these TRAPase‐positive cells have been previously documented [8, 12]. Briefly, 5 × 10 4 viable cells/ml were cultured in 15 × 10 mm Linbro wells (0.5 ml/well) (Flow Laboratories; McLean, VA) with supplemented medium 199 (Whittaker BioProduct; Walkersville, MO) containing 20% FCS, 0.3% agar, and optimal concentrations of CESJ medium as a source of O‐CSF (see below).…”

Section: Methodsmentioning

confidence: 99%

Conditions that Support Long‐Term Production of Osteoclast Progenitors In Vitro

et al. 1997

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To better understand the mechanisms of osteoclast precursor development from hematopoietic stem cells, we examined the conditions that support the production of osteoclast progenitors, osteoclast colony-forming units (CFU-O), from long-term bone marrow cultures established under myeloid (Dexter's) and lymphoid (Whitlock and Witte's) conditions. Nonadherent cells harvested weekly from myeloid or lymphoid longterm cultures were assayed for CFU-O-derived colony formation in agar in the presence of a murine osteoclast colony-stimulating factor. The myeloid system supported CFU-O production for weeks, but the system produced many other types of myeloid colonies and cells as well, and quantification of CFU-O-derived colonies was difficult. The lymphoid long-term culture system also produced CFU-O; however, CFU-O production in the lymphoid system appeared more selective than in the myeloid system, but was transient. Interestingly, the addition of medium containing G-CSF to these cultures greatly enhanced (>200%) the CFU-O production. This enhanced CFU-O production was confirmed using bone marrow cultures established on a defined marrow stromal cell line under lymphoid conditions and supplemented with recombinant murine G-CSF. Thus, G-CSF facilitates the development of clonogenic osteoclast progenitors from hematopoietic stem cells in lymphoid long-term culture conditions. This culture system may serve as a useful model for ex vivo generation of osteoclast progenitors.

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Section: Methodsmentioning

confidence: 99%