Martinez-Corral Ines, Makinen Taija
Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
Methods Mol Biol. 2018;1846:37-53. doi: 10.1007/978-1-4939-8712-2_3.
Lineage tracing allows for identification of all progeny produced by a single cell or groups of cells and can thus be used to assess developmental fate of cells. Here we focus on one of the most widely used lineage tracing approaches that utilize the Cre/loxP system for site-specific genetic recombination in studying the developmental origins of lymphatic endothelial cells (LECs) in the mouse embryo. We discuss general considerations for a successful Cre/loxP based lineage tracing experiment and provide information about strains that are available for genetic lineage tracing of LECs. A protocol for lineage tracing analysis of the lymphatic vasculature by whole-mount immunofluorescence in two embryonic tissues, the skin and the mesentery, is also provided.
谱系追踪能够识别单个细胞或细胞群体产生的所有后代,因此可用于评估细胞的发育命运。在这里,我们重点关注一种应用最为广泛的谱系追踪方法,该方法利用Cre/loxP系统进行位点特异性基因重组,以研究小鼠胚胎中淋巴管内皮细胞(LEC)的发育起源。我们讨论了基于Cre/loxP的成功谱系追踪实验的一般注意事项,并提供了可用于LEC遗传谱系追踪的品系信息。此外,还提供了一种通过全组织免疫荧光对皮肤和肠系膜这两种胚胎组织中的淋巴管系统进行谱系追踪分析的方案。