Lineage tracing in the intestinal epithelium.
Lineage tracing in the intestinal epithelium.
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DOI：
10.1002/9780470151808.sc05a04s13
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发表时间：
2010-05-01
影响因子：
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通讯作者：
Clevers, Hans
中科院分区：
文献类型：
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作者：
Barker, Nick;Clevers, Hans
本单元描述了使用 Lgr5-EGFP-ires-CreERT2/Rosa26lacZ 小鼠模型对 Lgr5(+ve) 肠干细胞进行体内谱系追踪的理论和详细方案。通过给予限制剂量的激素他莫昔芬，可以在任何年龄的小鼠中启动谱系追踪。这会激活 Lgr5(+ve) 干细胞中的 lacZ 报告基因，随后当它们在正常体内平衡期间重新填充上皮时，将这种永久遗传标记传递给它们的后代。由于 Lgr5(+ve) 细胞是长寿、自我更新的干细胞，因此它们不断产生 lacZ 后代，这有助于小鼠整个生命周期的组织更新。相同的方案可用于对其他 Lgr5(+ve) 干细胞群（包括毛囊和胃中的干细胞群）进行体内谱系追踪。
This unit describes the theory and detailed protocols for performing in vivo lineage tracing from Lgr5(+ve) intestinal stem cells using an Lgr5-EGFP-ires-CreERT2/Rosa26lacZ mouse model. Lineage tracing can be initiated in mice at any age by administering limiting doses of the hormone tamoxifen. This activates the lacZ reporter gene in the Lgr5(+ve) stem cells, which subsequently transmit this permanent genetic mark to their progeny as they repopulate the epithelium during normal homeostasis. Because the Lgr5(+ve) cells are long-lived, self-renewing stem cells, they continuously generate lacZ progeny, which contribute to tissue renewal over the entire lifetime of the mouse. The same protocols can be applied to performing in vivo lineage tracing from other Lgr5(+ve) stem cell populations, including those in the hair-follicle and stomach.