在诱导多能干细胞干细胞衍生的心肌细胞中记录和解释活性过渡物
Maedeh Mozneb1,2,3,4, Jemima Moses1,2,3,4,5, Madelyn Arzt1,2,3,4
1Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, California.
Current protocols
|October 24, 2024
概括
这项研究引入了一种使用人类诱导多能干细胞衍生心肌细胞 (hiPSC-CMs) 中遗传编码指标 (GCaMP) 的标准化方法,以准确测量过渡物. 这些协议确保了用于心肌细胞电生理学研究的可靠数据处理和分析.
科学领域:
- 心血管科学 心血管科学
- 细胞电生理学 细胞电生理学
- 生物医学成像技术 生物医学成像技术
背景情况:
- 信号传递对于心肌细胞激发-收缩合和节律收缩至关重要.
- 目前用于追踪细胞内的现有方法缺乏标准化,导致信号处理偏差和解释挑战.
- 准确地解释短暂信号对于理解心肌细胞电生理学至关重要.
研究的目的:
- 建立一个标准化的方法来记录和分析心肌细胞中的过渡.
- 在人类诱导的多能干细胞干细胞衍生的心肌细胞 (hiPSC-CMs) 中利用基因编码的指标 (GCaMP) 进行精确的信号检测.
- 为数据提取,处理和可视化提供协议,尽量减少信号处理偏差.
主要方法:
- 采用基因编码指标 (GCaMP) 的人类诱导多能干细胞衍生心肌细胞 (hiPSC-CMs) 进行基于光的记录.
- 使用 ImageJ 和 MATLAB 软件来提取和处理的短暂波形.
- 开发hiPSC维护,扩展,差异化,视频录制和信号分析的协议.
- 实施一个支持协议,用于将hiPSC-CMs复制到多种不同培养平台上,以实现数据复杂化.
主要成果:
- 展示了维护,扩展和区分GCaMP高PSC的综合方法.
- 概述了用于捕获GCaMP hiPSC-CMs中的过渡的视频录制技术.
- 提供了详细的信号提取,预处理,分析和可视化瞬态数据的方法.
- 根据对心肌细胞功能的生理相关性,促进了波形特征的分类.
结论:
- 提出的方案为研究心肌细胞中的动态提供了一种标准化和可靠的方法.
- 这种方法可以准确地解释的短暂信号,这对于推进心血管科学至关重要.
- 该方法支持数据复杂化,允许在各种平台上进行心肌细胞培养,从而增强研究能力.
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