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亲水支架细胞培养状态检测的染色方法

发表时间:2017-09-15  浏览量:884  下载量:102
全部作者: 皮彦斌,孟庆阳,敖英芳
作者单位: 北京大学第三医院运动医学研究所
摘 要: 目的:提出一种检测亲水支架上细胞培养状态的方法。方法:将待检测细胞接种到亲水支架上进行培养,至待检测细胞在亲水支架上的生长融合率为90%~95%时,获得支架-细胞复合体系。采用吖啶橙染色液对支架-细胞复合体系进行荧光染色,并采用PBS对染色后的支架-细胞复合体系进行洗涤。将洗涤后的支架-细胞复合体系置于激光扫描共聚焦显微镜下,利用绿色荧光通道和橙色荧光通道进行观察,从而检测得到亲水支架上细胞的培养状态。结果:通过这种方法,激光扫描共聚焦显微镜下的细胞分布情况显示十分清晰,不存在亲水支架对荧光染色液的吸附所造成的干扰,使细胞分布及生长状态的分析更加直观、准确。结论:本研究提供的方法能够清晰、准确地检测亲水支架上细胞的分布情况及生长状态,可以对亲水支架上的细胞进行无损、实时、动态检测,且操作简便,便于规模化推广应用。
关 键 词: 生物医学工程学;吖啶橙染色液;激光扫描共聚焦显微镜;亲水支架
Title: A staining method to detect cellular culture state on hydrophilic scaffold
Author: PI Yanbin, MENG Qingyang, AO Yingfang
Organization: Institute of Sports Medicine, Peking University Third Hospital
Abstract: Objective: In present study, a method was introduced to detect the state of cells cultured on the hydrophilic scaffold. Methods: Stem cells were cultured on a hydrophilic scaffold to build a cell-scaffold complex with a confluence of 90%-95%. The fluorescence staining of cell-scaffold complex was produced by acridine orange staining, and rinsed by PBS to erase the overdoes acridine orange dyes. After fluorescent staining, the cell-scaffold complex was observed under green and orange channels of laser scanning confocal microscope. Results: By this method, the state and distribution of cultured cells could be displayed clearly and precisely under laser scanning confocal microscope, without the interference of unintended staining on hydrophilic scaffold. Conclusion: The present study provides a harmless and simple method to detect the distributions and growth states of cells cultured on the hydrophilic scaffold, and it is suitable for dynamic monitoring of the states of cultured cells, and useful for wild-spread.
Key words: biomedical engineering; acridine orange dyes; laser scanning confocal microscope; hydrophilic scaffold
发表期数: 2017年9月第17期
引用格式: 皮彦斌,孟庆阳,敖英芳. 亲水支架细胞培养状态检测的染色方法[J]. 中国科技论文在线精品论文,2017,10(17):1927-1933.
 
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