使用流式细胞计量对差异化髓管的光表征
Andy Nolan1,2, Robert A Heaton2, Petra Adamova3
1Research Institute for Sport and Exercise Sciences, Liverpool John Moores University, Liverpool, UK.
概括
流式细胞计可以成功地分析骨肌肉的原始细胞 (神经质细胞) 和分化的神经管. 线粒体质量,活性氧物种和DNA含量的关键差异区分了髓管,尽管激光散射检测到的尺寸相似.
科学领域:
- 细胞生物学 细胞生物学
- 骨肌肉研究 骨肌肉研究
- 流细胞计的应用 流细胞计的应用
背景情况:
- 流细胞计是评估髓细胞体群体的标准.
- 缺乏使用流细胞计学对差异化肌管进行综合性关门策略.
- 与髓细胞相比,肌管表现出显著的代谢和蛋白质变化.
研究的目的:
- 确定流细胞计用于分析肌细胞体和肌管的实用性.
- 描述神经细胞和神经管之间的形态和代谢差异.
- 在异质细胞悬浮体内开发用于肌管识别的封闭策略.
主要方法:
- 使用流细胞计,分析了C2C12小鼠细胞的形态和代谢重编程.
- 激光散射 (FSC/SSC) 测量了细胞形态;光探针量化了线粒体质量,ROS和DNA含量.
- 肌重链 (MyHC) 免疫类型鉴定证实了分化;使用Annexin V/propidium iodide评估了生存能力.
主要成果:
- 与线粒体相比,线粒体质量 (159%),ROS产量 (303%),DNA含量 (18%) 和MyHC表达率 (147%) 显著提高.
- 激光散射表明神经细胞和神经管的尺寸相似,但"卷起来的"神经管形态改变了散射特性.
- 肌管表现出更大的异质性,确定了两个不同的子群体,一个是可行的,另一个可能含有亡体.
结论:
- 流细胞计对于分析肌管是有效的,肌管质量增加,ROS和DNA含量是与MyHC表达相关的关键标识符.
- 在分析过程中肌管"滚动"使得通过激光散射来确定尺寸变得复杂,需要使用替代的识别策略.
- 声学聚焦可能优于水力动力聚焦,因为它们的异质性,可以解决不同的神经管子群.
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