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从正常小梁骨表面分离的成骨细胞的培养与行为

Culture and behavior of osteoblastic cells isolated from normal trabecular bone surfaces.

作者信息

Marie P J, Lomri A, Sabbagh A, Basle M

机构信息

INSERM Unité 18, Hôpital Lariboisière, Paris, France.

出版信息

In Vitro Cell Dev Biol. 1989 Apr;25(4):373-80. doi: 10.1007/BF02624601.

DOI:10.1007/BF02624601
PMID:2541129
Abstract

We report the characterization of human osteoblastic cells that were derived from the surface of trabecular bone fragments. After removal of bone marrow cells, the bone lining osteoblastic cells lining the bone surface were obtained by migration and proliferation from the trabecular surface onto a nylon mesh. The isolated population proliferated in culture and exhibited osteoblastic phenotype. Cultured cells show a regular arrangement in vitro and exhibited multiple interconnecting junctions on scanning electron microscopic examination. Immunocytochemical staining showed that the cells produced almost exclusively type I collagen. Bone-surface-derived cells responded to 1-34 human parathyroid hormone by increasing intracellular cyclic AMP. Cell cultures exhibited high alkaline phosphatase activity, which was unaffected by 1,25 (OH)2 vitamin D. Untreated cells produced high levels of osteocalcin, a bone-specific protein, and they responded to 1,25(OH) vitamin D by increasing osteocalcin synthesis in a dose-dependent manner. Although cells cultured for up to 5 mo. still produced osteocalcin, the response to 1,25(OH)2D decreased after multiple passages. This study shows that the bone cell populations isolated from trabecular bone surfaces are enriched in osteoblast precursors and mature osteoblastic cells.

摘要

我们报告了源自小梁骨碎片表面的人成骨细胞的特性。去除骨髓细胞后,通过从骨小梁表面迁移和增殖到尼龙网上,获得了覆盖在骨表面的骨衬里成骨细胞。分离出的细胞群体在培养中增殖并表现出成骨细胞表型。培养的细胞在体外呈规则排列,扫描电子显微镜检查显示有多个相互连接的连接点。免疫细胞化学染色表明,这些细胞几乎只产生I型胶原蛋白。骨表面来源的细胞通过增加细胞内环磷酸腺苷对1-34人甲状旁腺激素作出反应。细胞培养物表现出高碱性磷酸酶活性,这不受1,25(OH)2维生素D的影响。未处理的细胞产生高水平的骨钙素,一种骨特异性蛋白,并且它们通过以剂量依赖的方式增加骨钙素的合成对1,25(OH)维生素D作出反应。尽管培养长达5个月的细胞仍产生骨钙素,但多次传代后对1,25(OH)2D的反应降低。这项研究表明,从小梁骨表面分离的骨细胞群体富含成骨细胞前体和成熟的成骨细胞。

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