| 小鼠气管平滑肌原代细胞的分离、培养与鉴定 |
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| 引用本文： | 叶致豪,许文豪,刘庆华,沈金花,彭勇波. 小鼠气管平滑肌原代细胞的分离、培养与鉴定[J]. 实验动物与比较医学, 2016, 24(4): 364-368 |
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| 作者姓名： | 叶致豪 许文豪 刘庆华 沈金花 彭勇波 |
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| 作者单位： | 中南民族大学,中南民族大学,中南民族大学,中南民族大学,中南民族大学 |
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| 基金项目： | 国家自然科学基金项目（30900816, 31371307），湖北省自然科学基金项目（2014CFC116） |
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| 摘 要： | 目的：气管平滑肌细胞的异常增殖和收缩是引起气道重塑和哮喘发病的重要原因之一，本研究的目的在于建立一种可靠的通过组织块贴壁法分离培养原代小鼠气管平滑肌细胞及免疫组化鉴定的方法。方法：体式显微镜下立体分离小鼠气管平滑肌组织，组织块贴壁法培养原代细胞，对分离培养细胞通过免疫组化方法进行鉴定，并用MTT法对其增殖特性进行检测。结果：从BALB/c雄性小鼠分离气管平滑肌组织，剪碎为1mm3，用含1%青-链霉素的PBS及培养液漂洗，使组织块贴于培养皿底，并加入5mL培养液，放入37℃、5 % CO2的细胞培养箱培养，3-5天后有明显梭状细胞从组织块爬出，5-6天后，细胞可见明显“峰-谷”结构。经免疫荧光鉴定，在传代、纯化后，可得纯度为99%以上的气管平滑肌细胞。用MTT法测量其生长曲线。结论：本方法操作简单、经济，获得的气管平滑肌细胞具有较好增殖能力，细胞数量和纯度能够满足后续细胞生物学实验研究的需要。
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| 关 键 词： | 气管平滑肌细胞；原代培养；组织块贴壁法；免疫荧光；MTT法 |
| 收稿时间： | 2016-01-25 |
| 修稿时间： | 2016-02-25 |
Isolation, culture and identification of primary airway smooth muscle cells from mice |
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| YE Zhi-hao,XU Wen-hao,LIU Qing-hu,SHEN Jin-hua and PENG Yong-bo. Isolation, culture and identification of primary airway smooth muscle cells from mice[J]. Laboratory Animal and Comparative Medicine, 2016, 24(4): 364-368 |
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| Authors: | YE Zhi-hao XU Wen-hao LIU Qing-hu SHEN Jin-hua PENG Yong-bo |
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| Affiliation: | Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, Institute of Medical Biology, School of Life Science, South-Central University for Nationalities, Wuhan 430074, China;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, Institute of Medical Biology, School of Life Science, South-Central University for Nationalities, Wuhan 430074, China;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, Institute of Medical Biology, School of Life Science, South-Central University for Nationalities, Wuhan 430074, China;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, Institute of Medical Biology, School of Life Science, South-Central University for Nationalities, Wuhan 430074, China;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, Institute of Medical Biology, School of Life Science, South-Central University for Nationalities, Wuhan 430074, China |
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| Abstract: | |
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| Keywords: | ASMCs Primary culture Tissue section adherence Immunofluorescence MTT method |
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