Liu Xiuxiu, Weng Wendong, He Lingjuan, Zhou Bin
State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.
School of Life Sciences, Westlake University, Hangzhou, China.
Nat Protoc. 2023 Jul;18(7):2349-2373. doi: 10.1038/s41596-023-00833-8. Epub 2023 Jun 2.
The ability to experimentally measure cell proliferation is the basis for understanding the sources of cells that drive organ development, tissue regeneration and repair. Recently, we generated a genetic approach to detect cell proliferation: we used genetic lineage-tracing technologies to achieve seamless recording of in vivo cell proliferation in a tissue-specific manner. We provide a detailed protocol (generation of mouse lines, characterization of mouse lines, mouse line crossing and cell-proliferation tracing) for using this genetic system to study cell proliferation. This cell-proliferation tracing system, which we term 'ProTracer' (Proliferation Tracer), permits lifelong noninvasive monitoring of cell proliferation of specific cell lineages in live animals. Compared with other short-term strategies that require execution of animals, ProTracer does not require sampling or animal sacrifice for tissue processing. To highlight these features, we used ProTracer to study the proliferation of hepatocytes during liver homeostasis and after tissue injury in mice. We show that the protocol is applicable to study any in vivo cell proliferation, which takes ~9 months to finish from mouse generation to data analysis. This protocol can easily be carried out by researchers skilled in mouse-related experiments.
通过实验测量细胞增殖的能力是理解驱动器官发育、组织再生和修复的细胞来源的基础。最近，我们开发了一种检测细胞增殖的遗传学方法：我们利用遗传谱系追踪技术以组织特异性方式实现对体内细胞增殖的无缝记录。我们提供了一个详细的方案（小鼠品系的生成、小鼠品系的表征、小鼠品系杂交和细胞增殖追踪），用于使用这种遗传系统研究细胞增殖。我们将这个细胞增殖追踪系统称为“ProTracer”（增殖追踪器），它允许对活体动物中特定细胞谱系的细胞增殖进行终身无创监测。与其他需要处死动物的短期策略相比，ProTracer不需要取样或牺牲动物来进行组织处理。为了突出这些特点，我们使用ProTracer研究了小鼠肝脏稳态期间和组织损伤后肝细胞的增殖。我们表明该方案适用于研究任何体内细胞增殖，从小鼠生成到数据分析大约需要9个月时间。该方案可以由精通小鼠相关实验的研究人员轻松完成。
