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一个体外平台,用于量化细胞周期阶段长度在人类初级肠干细胞

Michael J Cotton¹, Pablo Ariel², Kaiwen Chen¹

¹Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, North Carolina.

bioRxiv : the preprint server for biology

|October 24, 2023

概括

研究人员开发了新的平台来分析活人肠干细胞 (ISC) 中的细胞周期阶段. 这些工具可以详细研究ISC细胞周期调节和对药物剂的反应.

科学领域:

干细胞生物学干细胞生物学
细胞周期调节细胞周期调节
胃肠道研究胃肠道研究

背景情况:

肠上皮具有动态控制细胞周期阶段长度的作用.
没有现有的实验平台可以直接分析活人肠干细胞 (ISC) 中的细胞周期阶段.

研究的目的:

开发新的实验平台,用于分析活体初级人类ISC中的细胞周期阶段.
为了使ISC细胞周期调节和药物反应的详细研究.

主要方法:

用于支持ISC生长和共聚焦成像的室内幻灯片的原体压缩机的3D打印.
在人类ISC中利用了PIP-FUCCI和PIP-H2A光细胞循环记者结构.
使用实时成像,EDU脉冲和CellPose2和TrackMate的计算分析管道.

主要成果:

开发了支持ISC循环和共聚焦显微镜的室内幻灯片.
PIP-FUCCI ISC对于快照细胞周期阶段分配是最优的.
通过实时成像,PIP-H2A ISC 便于通过实时成像进行自动化细胞周期阶段长度量化.

结论:

介绍了两种全面的平台,用于分析活体初级人类ISC的细胞周期阶段.
PIP-FUCCI和PIP-H2A结构允许在ISC中进行细胞循环分析的不同方法.
这些平台对于未来的药物效应,细胞循环调节和ISC生物学研究具有价值.

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