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In vitro osteoclast generation from different bone marrow fractions, including a highly enriched haematopoietic stem cell population

Naturevolume 321pages7981 (1986) | Download Citation



It is well established that the osteoclast is formed by fusion of post-mitotic, mononuclear precursors1 derived from circulating progenitor cells2. However, the precise haematopoietic origin of the osteoclast is unknown. We have investigated this here by fractionating mouse bone marrow and isolating haematopoietic stem cells using a three-step method combining equilibrium density centrifugation and two fluorescence-activated cell sortings (FACS)3, and have tested the ability of each bone marrow fraction, including highly purified haematopoietic stem cells, to generate osteoclasts during co-culture with preosteoclast-free embryonic long bones4,5. The osteoclast-forming capacity was found to increase with increasing stem cell purity. On the other hand, the culture time needed for osteoclast formation also increased with purification, suggesting the presence of progressively more immature progenitor cells. The pluripotent haematopoietic stem cell fractions with the highest purity needed preincubation with a stem cell-activating factor (interleukin-3) to activate the predominantly quiescent stem cells in vitro.

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  1. Laboratory of Cell Biology and Histology, University of Leiden, Rijnsburgerweg 10, 2333 AA, Leiden, The Netherlands

    • Ben A. A. Scheven
    •  & Peter J. Nijweide
  2. Radiobiological Institute TNO, Lange Kleiweg 151, 2288 GJ, Rijswijk, The Netherlands

    • Jan W. M. Visser


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