ISOLATED OSTEOCLASTS AND THEIR PRESUMED PROGENITOR CELLS, THE MONOCYTE, IN CULTURE
ISOLATED OSTEOCLASTS AND THEIR PRESUMED PROGENITOR CELLS, THE MONOCYTE, IN CULTURE
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DOI：
10.1002/jez.1402240306
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发表时间：
1982-01-01
影响因子：
--
通讯作者：
CAPLAN, AI
中科院分区：
文献类型：
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作者：
OSDOBY, P;MARTINI, MC;CAPLAN, AI
通过轻度胰蛋白酶消化从第 19 天胚胎鸡胫骨的骨内膜表面分离破骨细胞。破骨细胞富集是通过使细胞悬浮液通过选择性尺寸的 Nitex 筛选来实现的，包括最终将破骨细胞选择性保留在 12 μm 聚碳酸酯过滤器上，或通过 Nitex 筛选器顺序筛选并在 Percoll 梯度上分级。根据形态学标准，富集程序产生了 50-75% 的破骨细胞群体，而后一种分离方法为群体提供了较少的基质碎片。光学显微镜、透射和扫描电子显微镜观察结果表明，破骨细胞可以在培养物中维持长达10天，并保留破骨细胞的形态。这种形态包括特殊的褶皱质膜、大量线粒体、溶酶体和多核细胞质。酸性磷酸酶和丁酸酯酶组织化学测量支持这些形态学观察。通过Ficoll-hypaque梯度离心分离和纯化小鸡孵化的循环单核细胞，随后粘附到玻璃培养皿上。随着培养时间的推移，这些细胞形成多核细胞，但缺乏分离的破骨细胞的超微结构复杂性。描述了研究破骨细胞功能的独特培养系统。说明了该系统与单核细胞到巨细胞培养系统的相似之处和不同之处。
Osteoclasts were isolated from the endosteal surface of day-19 embryonic chick tibias by mild trypsinization. Osteoclast enrichment was achieved by passing cell supensions through Nitex screening of selective sizes, including the eventual selective retention of osteoclasts on 12 .mu.m polycarbonate filters or by sequential sieving through Nitex screens and fractionation on Percoll gradients. The enrichment procedures produced osteoclast populations of 50-75% based on morphological criteria with the latter isolation method providing populations with less matrix debris. The results of light microscopy, transmission and scanning electron microscopic observations indicate that osteoclasts can be maintained in culture for up to 10 days with retention of osteoclast morphology. This morphology includes a specialized ruffled plasma membrane, large numbers of mitochondria, lysosomes and a multinucleated cytoplasm. Acid phosphatase and butyrate esterase histochemical measurements support these morphological observations. Chick hatchling circulating monocytes were isolated and purified by Ficoll-hypaque gradient centrifugation with subsequent adhesion to glass petri dishes. With time in culture, these cells form multinucleated cells, but lack the ultrastructural complexity of the isolated osteoclasts. A unique culture system to study osteoclast function is described. Similarities and differences of this system to the monocyte-to-giant cell culture system are illustrated.