Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Copenhagen, Denmark.
Cell Plasticity and Regeneration Group, Regenerative Medicine Program, Institut d'Investigació Biomèdica de Bellvitge - IDIBELL, L'Hospitalet de Llobregat, Llobregat, Spain.
Methods Mol Biol. 2021;2258:29-40. doi: 10.1007/978-1-0716-1174-6_3.
Lineage-tracing experiments aim to identify and track the progeny and/or fate of cells. The use of inducible recombinases and fluorescent reporters has been instrumental in defining cellular hierarchies and allowing for the identification of stem cells in an unperturbed in vivo setting. The refinement of these approaches, labeling single cells, and the subsequent quantitative analysis of the clonal dynamics have allowed the comparison of different stem cell populations as well as establishing different mechanisms of cellular replenishment during steady-state homeostasis as well as during morphogenesis and disease. Utilizing this approach, it is now possible to establish the cellular hierarchy in a given tissue and the frequency of cell fate decisions on a population basis, thus providing a comprehensive analysis of cellular behavior in vivo. Although in this chapter we describe a protocol for lineage tracing of cells from fetal intestinal epithelium to the adult intestine, this approach can be widely applied to quantitatively assess the cell fate of any fetal cell during morphogenesis.
谱系追踪实验旨在鉴定和追踪细胞的后代和/或命运。诱导型重组酶和荧光报告基因的使用对于定义细胞层次结构以及在未受干扰的体内环境中鉴定干细胞非常重要。这些方法的改进，包括标记单个细胞以及随后对克隆动力学的定量分析，允许比较不同的干细胞群体，并确定在稳态平衡、形态发生和疾病过程中细胞补充的不同机制。利用这种方法，现在可以在给定组织中建立细胞层次结构，并基于群体对细胞命运决定的频率进行分析，从而对体内细胞行为进行全面分析。尽管在本章中我们描述了从胎儿肠上皮到成年肠谱系追踪细胞的方案，但这种方法可以广泛应用于定量评估形态发生过程中任何胎儿细胞的命运。
