通过使用流细胞计分离癌细胞亚群的代谢活性协议
Wiktoria Blaszczak1, Bobby White1, Pawel Swietach1
1Department of Physiology, Anatomy and Genetics, University of Oxford, Parks Road, OX1 3PT Oxford, UK.
STAR protocols
|June 2, 2024
概括
这项研究介绍了一种光激活细胞分类 (FACS) 方法,以基于代谢率来分离细胞. 该协议使用乳酸流出的pH值变化来识别用于细胞分类的高代谢细胞.
科学领域:
- 细胞生物学 细胞生物学
- 生物化学 生物化学
- 生物技术是生物技术.
背景情况:
- 细胞代谢异质性影响生物过程.
- 现有的方法缺乏通过代谢率有效分离细胞.
研究的目的:
- 开发一种光激活细胞分类 (FACS) 协议,根据代谢活动分离细胞.
- 为了使代谢上不同的亚种群的分离.
主要方法:
- 光激活细胞分类 (FACS) 的适应用于细胞分离.
- 诱导乳酸流出,以创建可测量的pH短暂.
- 使用pH敏感染料通过pH特征检测代谢速率.
- 采用度测定方法来测量氧气消耗和酸的产生.
主要成果:
- 根据使用FACS的代谢率成功分离了细胞.
- 证明了乳酸流量,pH标志和代谢率之间的相关性.
- 通过氧气消耗和酸生产测定,通过排序的子群体之间确认了代谢对比.
结论:
- 开发的FACS协议有效地隔离了具有不同代谢速率的细胞.
- 这种方法为研究细胞代谢异质性提供了强大的工具.
- 能够进一步研究代谢上不同的细胞群的功能作用.
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