Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA, USA.
Methods Mol Biol. 2022;2544:207-216. doi: 10.1007/978-1-0716-2557-6_15.
A fundamental part of understanding how a tissue regenerates is how individual cells that make up the tissue divide. This protocol describes a method to label a random and representative sampling of hepatocytes throughout the mouse liver and quantify their cell division and clonal expansion during homeostasis and after liver injury. Clonal expansion is quantified through 3D imaging, which has improved accuracy over 2D imaging. This information can be used to answer questions such as which hepatocytes contribute to hepatocyte renewal during homeostasis and during regeneration after injury, as well as how much individual hepatocytes in the different zones of the liver divide.
理解组织如何再生的一个基本部分是组成组织的单个细胞如何分裂。本方案描述了一种标记小鼠肝脏中随机和代表性的肝细胞样本的方法,并在肝内稳态和肝损伤后定量分析其细胞分裂和克隆扩增。通过 3D 成像对克隆扩增进行定量,其准确性优于 2D 成像。这些信息可用于回答以下问题,例如在肝内稳态和损伤后再生期间,哪些肝细胞有助于肝细胞更新,以及肝脏不同区域的单个肝细胞分裂的程度。