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首页> 外文期刊>Journal of Cellular Physiology >Human trabecular bone-derived osteoblasts support human osteoclast formation in vitro in a defined, serum-free medium.
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Human trabecular bone-derived osteoblasts support human osteoclast formation in vitro in a defined, serum-free medium.

机译：人类小梁bone-derived成骨细胞的支持定义人类体外破骨细胞的形成,无血清培养基。

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While it has been assumed that osteoblasts in the human support osteoclast formation, in vitro evidence of this is currently lacking. We tested the ability of normal human trabecular bone-derived osteoblasts (NHBCs) to support osteoclast formation from human peripheral blood mononuclear cells (PBMC) in response to treatment with either 1alpha,25-dihydroxyvitamin D3 (1,25D) or parathyroid hormone (PTH), using a serum-replete medium previously used to support human osteoclast formation on a stroma of murine ST-2 cells. Under these conditions, NHBC did not support osteoclast formation, as assessed by morphological, histochemical, and functional criteria, despite our previous results demonstrating a link between induction of RANKL mRNA expression and NHBC phenotype in these media. We next tested a defined, serum-free medium (SDM) on NHBC phenotype, their expression of RANKL and OPG, and their ability to support osteoclast formation. SDM, containing dexamethasone (DEX) and 1,25D, induced phenotypic maturation of NHBC, based on the expression of STRO-1 and the bone/liver/kidney isoform of alkaline phosphatase (AP). PTH as a single factor did not induce phenotypic change. 1,25D and DEX induced the greatest ratio of RANKL:OPG mRNA, predictive of supporting osteoclast formation. Consistent with this, co-culture of NHBC with CD14+ PBMC, or bone marrow mononuclear cell (BMMC), or CD34+ BMMC precursors in SDM + 1,25D + DEX, resulted in functional osteoclast formation. Osteoclast formation also occurred in PTH + DEX stimulated co-cultures. Interestingly, SDM supplemented with recombinant RANKL (25-100 ng/ml) and M-CSF (25 ng/ml), did not induce osteoclast formation from any of the osteoclast precursor populations in stromal-free cultures, unlike serum-replete medium. This study demonstrates that under the appropriate conditions, adult human primary osteoblasts can support de novo osteoclast formation, and this model will enable the detailed study of the role of both cell types in this process.
机译：虽然一直认为的成骨细胞人类的支持形成破骨细胞,在体外目前缺乏这方面的证据。正常的人类小梁的能力bone-derived成骨细胞(NHBCs)的支持从人类外周血破骨细胞的形成单核细胞(PBMC)对治疗的反应用1α,25-dihydroxyvitamin D3 (1,25 d)或甲状旁腺激素(甲状旁腺素),使用一个serum-replete介质之前用于支持人类的破骨细胞形成的小鼠基质四白细胞。支持破骨细胞的形成,如评估形态学、组织化学和功能标准,尽管我们以前的结果展示感应RANKL的之间的联系信使rna表达和NHBC表型在这些媒体。介质(SDM) NHBC表型,他们的表情RANKL和功能,他们支持的能力破骨细胞的形成。地塞米松(DEX)和1,25 d,诱导表型NHBC成熟,基于的表达STRO-1和骨/肝/肾同种型的碱性磷酸酶(美联社)。没有引起表型变化。诱导RANKL的最大比率:功能mRNA,预测支持破骨细胞的形成。与此一致的是,共培养的NHBCCD14 + PBMC,或骨髓单核细胞敏捷,导致功能破骨细胞的形成。破骨细胞的形成也发生在甲状旁腺素+敏捷刺激培养。补充了重组RANKL (25 - 100ng / ml)和csf (25 ng / ml),没有诱导破骨细胞形成的破骨细胞前体stromal-free人口文化,不像serum-replete媒介。表明在适当的条件下,成人主要造骨细胞支持新创破骨细胞的形成,这模型将使角色的详细研究在这个过程中两种细胞类型。

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