单细胞分类来自人类脱皮的叶子牙的免疫表型介质干细胞
Ayona Gupta1, Risani Mukhopadhyay1, Himanshi Khandelwal1
1Manipal Institute of Regenerative Medicine, Bengaluru, Manipal Academy of Higher Education, Manipal.
Journal of visualized experiments : JoVE
|November 27, 2023
概括
这项研究介绍了一种使用流细胞测量来隔离从落叶牙中纯干细胞的方法. 这种技术提高了细胞活力和增殖,用于再生医学应用.
科学领域:
- 再生生物学 再生生物学
- 细胞生物学 细胞生物学
- 免疫学 免疫学 免疫学
背景情况:
- 介质干细胞 (MSC) 具有广泛的分化潜力和免疫调节特性,对于再生生物学至关重要.
- 多种来源和体外条件导致的MSC细胞模两可,导致功能异质,阻碍治疗应用.
- 需要标准化方法来获得纯化,均的MSC种群,以获得安全有效的再生医学.
研究的目的:
- 开发和验证一项使用光辅助单细胞分类来识别和净化人类脱皮叶状牙 (SHED) 的干细胞的协议.
- 通过隔离特定的亚群来解决MSC的功能异质性,以治疗用途.
- 建立一种可靠的方法来获得可行的和繁殖的MSCs,用于下游应用,如谱系特异化.
主要方法:
- 使用多参数流细胞测量来分析SHEDs上的表面标记物表达 (CD90-FITC,CD73-PerCP-Cy5.5,CD105-APC,CD44-V450).
- 采用光辅助单细胞分类,定义基于特定正负标记表达的纳入标准,以隔离所需的MSC亚群.
- 在分类后评估细胞活力和增殖,以确保纯化干细胞群的质量.
主要成果:
- 通过同时表达CD90,CD73,CD105和CD44标记物来确定MSCs.
- 从通道7开始,观察到表达所有四个标记物的细胞百分比下降.
- 通过使用定义的流式细胞计量标准,成功排序了可行的和繁殖的MSC亚群,证明了隔离协议的有效性.
结论:
- 光辅助单细胞分类提供了一种强大的方法,用于从SHED中净化同质的MSC种群.
- 开发的协议确保了高细胞活力和增殖,使分类细胞适合再生医学和进一步研究.
- 这种方法可以适应其他单细胞系统,以改善隔离,并收集详细的细胞表面标记信息.
相关概念视频
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