MINERALIZED BONE NODULES FORMED INVITRO FROM ENZYMATICALLY RELEASED RAT CALVARIA CELL-POPULATIONS
MINERALIZED BONE NODULES FORMED INVITRO FROM ENZYMATICALLY RELEASED RAT CALVARIA CELL-POPULATIONS
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DOI：
10.1007/bf02556874
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发表时间：
1986-03-01
影响因子：
4.2
通讯作者：
ANTOSZ, ME
中科院分区：
文献类型：
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作者：
BELLOWS, CG;AUBIN, JE;ANTOSZ, ME
从连续酶消化的胎鼠颅骨获得的单细胞悬液在长期培养中培养，在抗坏血酸、β-甘油磷酸钠和地塞米松的存在下培养，以确定这些群体形成矿化骨的能力。在单独加入抗坏血酸和β-甘油磷酸或抗坏血酸的成骨细胞培养中，三维结节(.apprx.75微米厚)，融合后3天可检测到覆盖有类似成骨细胞的多边形细胞。再过3-4天，结节变得肉眼可见(直径可达3毫米)。只有在有机磷酸盐存在的情况下，它们才会矿化。在没有抗坏血酸的情况下，结节是不会形成的。地塞米松导致结节数量显著增加。组织学上，结节类似编织骨，覆盖结节的细胞碱性磷酸酶染色强烈。特异性抗体免疫标记显示，矿物质相关的I型胶原染色较强，III型胶原和骨结素染色较弱，II型胶原染色检测不到。结节不是从种群I发展而来的，种群II-V形成的结节数量与所培养细胞的数量呈线性关系(r=0.99)。结果表明，在抗坏血酸和有机磷的存在下，酶促释放的颅骨细胞在体外可以形成矿化的骨结节。
Single-cell suspensions obtained from sequential enzymatic digestions of fetal rat calvaria were grown in long-term culture in the presence of ascorbic acid, Na .beta.-glycerophosphate, and dexamethasone to determine the capacity of these populations to form mineralized bone. In cultures of osteoblastlike cells grown in the presence of ascorbic acid and .beta.-glycerophosphate or ascorbic acid alone, three-dimensional nodules (.apprx. 75 .mu.m thick) covered by polygonal cells resembling osteoblasts could be detected 3 days after confluency. The nodules became macroscopic (up to 3 mm in diameter) after a further 3-4 days. Only in the presence of organic phosphate did they mineralize. Nodules did not develop without ascorbic acid in the medium. Dexamethasone caused a significant increase in the number of nodules. Histologically, nodules resembled woven bone and the cells covering the nodules stained strongly for alkaline phosphatase. Immunolabelling with specific antibodies demonstrated intense staining for type I collagen that was mineral-associated, a weaker staining for type III collagen and osteonectin, and undetectable staining for type II collagen. Nodules did not develop from population I and the number of nodules formed by populations II-V bore a linear relationship to the number of cells plated (r = 0.99). The results indicate that enzymatically released calvaria cells can form mineralized bone nodules in vitro in the presence of ascorbic acid and organic phosphate.