| 胰酶消化法纯化诱导不同年龄段SD大鼠骨髓破骨细胞的体外培养 |
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| 引用本文： | 董强,梁星,陈悦,徐凌,张庆鸿,夏露,陈明,付俊. 胰酶消化法纯化诱导不同年龄段SD大鼠骨髓破骨细胞的体外培养[J]. 华西口腔医学杂志, 2006, 24(4): 350-352 |
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| 作者姓名： | 董强 梁星 陈悦 徐凌 张庆鸿 夏露 陈明 付俊 |
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| 作者单位： | 贵州医学院附属医院口腔科;四川大学华西口腔医院,修复科,四川,成都,610041;口腔生物医学工程教育部重点实验室,四川大学,四川,成都,610041 |
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| 基金项目： | 国家自然科学基金;教育部优秀青年教师资助计划;高等学校博士学科点专项科研项目 |
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| 摘 要： | 目的 用胰酶消化法纯化获取大量高纯度破骨细胞，为进行相关分子生物学研究奠定基础。方法 以1,25-（OH）2D3 /地塞米松诱导培养不同年龄段（1、12、24、36 d）SD大鼠的骨髓破骨细胞，采用0.25%胰蛋白酶/0.02%乙二胺四乙酸纯化。通过倒置相差显微镜观察细胞形态，采用抗酒石酸酸性磷酸酶（Trap）染色和扫描电镜观察鉴定破骨细胞，并计数分析。结果 1、12、24、36 d组大鼠骨髓破骨细胞均培养成功，经Trap染色和扫描电镜观察证实为体外有噬骨能力的破骨细胞，纯化率达到90％。1 d和12 d组破骨细胞出现较早且数量较多，细胞计数两组间无统计学差异（P>0.05），但它们与其余各组均有统计学差异（P<0.05）。结论 采用胰酶消化法纯化1,25-（OH）2D3 /地塞米松诱导培养的大鼠骨髓破骨细胞，纯度较高；选用大鼠以10-12 d龄较为适宜。
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| 关 键 词： | 破骨细胞 胰酶消化 培养 纯化 |
| 文章编号： | 1000-1182（2006）04-0350-03 |
| 收稿时间： | 2005-10-20 |
| 修稿时间： | 2005-10-202006-02-23 |
Isolation of Bone Marrow-derived Rat Osteoclast-like Cells with the Digestion of Trypsin |
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| DONG Qiang,LIANG Xing,CHEN Yue,XU Ling,ZHANG Qing-hong,XIA Lu,CHEN Ming,FU Jun. Isolation of Bone Marrow-derived Rat Osteoclast-like Cells with the Digestion of Trypsin[J]. West China journal of stomatology, 2006, 24(4): 350-352 |
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| Authors: | DONG Qiang LIANG Xing CHEN Yue XU Ling ZHANG Qing-hong XIA Lu CHEN Ming FU Jun |
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| Affiliation: | 1. Key. Laboratory of Oral Biomedical Engineering Ministry of Education, Sichuan University, Chengdu 610041, China; 2. Dept. of Prosthodontics, West China College of Stomatology, Sichuan University, Chengdu 610041, China |
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| Abstract: | OBJECTIVE: To obtain highly enriched osteoclasts in vitro. METHODS: The bone marrow cells of 1-day, 12-day, 24-day and 36-day old Sprague-Dawley (SD) rats were separately cultured with the presence of 1,25-(OH)2D3 and dexamethasone, and the osteoclast-like cells were identified by Trap staining and scanning electron microscope observation and were purified by trypsin digestion. The count of Trap positive osteoclast-like cells was analyzed statistically. RESULTS: The osteoclast-like cells of each group were Trap positive cells and could form the bone absorption lacunas in vitro. The amounts of osteoclast-like cells were different statistically between the groups (P < 0.05) except between the groups of 1-day old and 12-day old (P > 0.05). Highly enriched osteoclast-like cells were harvested by the digestion of 0.25% trypsin and 0.02% EDTA. CONCLUSION: It's indicated that a large amount of highly enriched osteoclast-like cells could be obtained through the culture of bone marrow cells of 10-day and 12-day old SD rats with 1,25-(OH)2D3 and dexamethasone and the digestion of trypsin/EDTA. |
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| Keywords: | osteoclast trypsin digestion culture purify |
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