Brief report: Musashi1-eGFP mice, a new tool for differential isolation of the intestinal stem cell populations
Brief report: Musashi1-eGFP mice, a new tool for differential isolation of the intestinal stem cell populations
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DOI：
10.1002/stem.1428
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发表时间：
2013-10-01
期刊：
影响因子：
5.2
通讯作者：
Plateroti, Michelina
中科院分区：
文献类型：
--
作者：
Cambuli, Francesca Maria;Rezza, Amelie;Plateroti, Michelina
由于隐窝干细胞的存在，肠道上皮在整个生命过程中迅速而持续地自我更新。在小肠中已经发现了两个不同的细胞池，它们的位置(“+4”或隐窝的底部)、特定标记的表达(Bmi1/mTert或Lgr5/Ascl2)以及细胞周期特征都不同。有趣的是，RNA结合蛋白Musashi1在这两个群体中都有表达，因此是两种干细胞类型的潜在标记。为了定位、分离和研究肠上皮内表达Musashi1的细胞，我们在7kb的Msi1启动子的控制下获得了表达GFP荧光蛋白的转基因小鼠。GFP在小肠和大肠隐窝中的表达模式与Musashi1完全重叠，验证了我们的模型。通过荧光激活的细胞分类、细胞和分子分析，我们发现GFP阳性的Msi1表达细胞分为两个主要的池，对应于Lgr5和mTert表达的干细胞。有趣的是，监测这两个分类种群的细胞周期活动表明，它们都在积极地循环，尽管证实了细胞周期长度的差异。总之，我们新的基于Musashi1表达的报告小鼠模型是分离和研究肠上皮干细胞的有用工具。此外，这些小鼠独一无二地能够在同一动物模型中同时研究两个肠道干细胞池。干细胞2013；31：2273-2278
The intestinal epithelium self-renews rapidly and continuously throughout life, due to the presence of crypt stem cells. Two pools of these cells have been identified in the small intestine, which differ in position ("+4" or the bottom of the crypts), expression of specific markers (Bmi1/mTert or Lgr5/Ascl2), and cell cycle characteristics. Interestingly, the RNA-binding protein Musashi1 is expressed in both populations and therefore a potential marker for both stem cell types. In order to locate, isolate, and study Musashi1-expressing cells within the intestinal epithelium, we generated transgenic mice expressing GFP fluorescent protein under the control of a 7-kb Msi1 promoter. The expression pattern of GFP in the intestinal crypts of both small and large intestines completely overlapped that of Musashi1, validating our model. By using fluorescence-activated cell sorting, cellular, and molecular analyses, we showed that GFP-positive Msi1-expressing cells are divided into two major pools corresponding to the Lgr5- and mTert-expressing stem cells. Interestingly, monitoring the cell cycle activity of the two sorted populations reveals that they are both actively cycling, although differences in cell cycle length were confirmed. Altogether, our new reporter mouse model based upon Musashi1 expression is a useful tool to isolate and study stem cells of the intestinal epithelium. Moreover, these mice uniquely enable the concomitant study of two pools of intestinal stem cells within the same animal model. Stem Cells 2013;31:2273-2278