| 大鼠肌源性干细胞的分离、纯化和培养 |
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| 引用本文： | 叶锦,靳风烁,陈锦,王鹏,梁培禾,聂志林,李黔生.大鼠肌源性干细胞的分离、纯化和培养[J].中国组织工程研究与临床康复,2010,14(14). |
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| 作者姓名： | 叶锦 靳风烁 陈锦 王鹏 梁培禾 聂志林 李黔生 |
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| 作者单位： | 解放军第三军医大学大坪医院野战外科研究所泌尿外科,重庆市,400042 |
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| 摘 要： | 背景:要获得满足临床需要的肌源性干细胞,体外筛选和扩增已成为关键环节.目的:拟建立稳定高效的成年大鼠肌源性干细胞的分离培养、纯化方法.方法:成年SD大鼠麻醉后,无菌条件下取骨骼肌,采用Ⅺ型胶原酶、Dispase和胰酶消化法获得肌源性干细胞,以密度梯度离心法和差速贴壁法进行纯化.记录细胞生长曲线,MTT比色法观察不同接种密度对细胞生长的影响,采用免疫细胞化学染色对细胞进行鉴定.结果与结论:原代肌源性干细胞体积较小,贴壁缓慢,折光性较好,多呈球形、梭形或纺锤形,增殖缓慢.传代培养后,加入含体积分数为20%血清浓度的完全培养基,以1×109L-1密度接种时活细胞数量最多,为适宜接种密度,传1～4代细胞生长状态良好.免疫细胞化学染色结果为desmin(+),CD34(+),CD45(-),Sea-1(+),证实通过体外原代培养获得了高纯度的肌源性干细胞,并成功扩增.
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| 关 键 词： | 酶 消化 密度梯度离心 差速贴壁 免疫细胞化学染色 分离培养 纯化 肌源性干细胞 |
Isolation,purification and cultivation of rat muscle-derived stem cells |
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| Ye Jin,Jin Feng-shuo,Chen Jin,Wang Peng,Liang Pei-he,Nie Zhi-lin,Li Qian-sheng.Isolation,purification and cultivation of rat muscle-derived stem cells[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2010,14(14). |
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| Authors: | Ye Jin Jin Feng-shuo Chen Jin Wang Peng Liang Pei-he Nie Zhi-lin Li Qian-sheng |
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| Institution: | Ye Jin,Jin Feng-shuo,Chen Jin,Wang Peng,Liang Pei-he,Nie Zhi-lin,Li Qian-sheng Department of Urology,Research Institute of Surgery,Daping Hospital,Third Military Medica University of Chinese PLA,Chongqing 400042,China |
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| Abstract: | BACKGROUND:In vitro screening and amplification are important links to harvest muscle-derived stem cells that are satisfactory to clinical requirement.OBJECTIVE:To probe into the method of isolation,culture and purification of skeletal muscle-derived stem cells from adult rats in vitro.METHODS:The skeletal muscle was obtained sterilely following adult Sprague Dawley rats were anesthetized.Muscle-derived stem cells were harvested using enzyme digestion with XI collagenase,Dispase and trypsogen,and then purif... |
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