Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium
Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium
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DOI：
10.1016/j.celrep.2013.09.005
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发表时间：
2013-10-01
期刊：
影响因子：
8.8
通讯作者：
Garcia, Marie-Isabelle
中科院分区：
文献类型：
--
作者：
Mustata, Roxana C.;Vasile, Gabriela;Garcia, Marie-Isabelle
使用最初开发用于培养成年肠上皮类器官的培养系统，从胎儿小鼠肠道中产生永生球体。从胎儿期到产后，球体比例逐渐减少，类器官的产量相应增加。与类器官一样，球状体表现出 Wnt 依赖性无限期自我更新特性，但表现出分化不良的表型，让人想起不完全尾部化的祖细胞。球体转录组与成体肠干细胞的转录组显着不同，Wnt 靶基因表达的重叠极小。受体 LGR4（而非 LGR5）对于它们的生长至关重要。 Trop2/Tacstd2 和 Cnx43/Gja1 这两个在球体中高度富集的标记物在整个胚胎第 14 天的肠上皮中表达。使用 Cnx43-CreER 和 Lgr5-CreERT2 小鼠进行子宫内和新生儿谱系追踪的比较，确定球状体生成细胞是参与产前肠上皮生成的发育祖细胞。在体外，球状细胞有可能分化成类器官，符合胎儿型肠道干细胞的资格。
Immortal spheroids were generated from fetal mouse intestine using the culture system initially developed to culture organoids from adult intestinal epithelium. Spheroid proportion progressively decreases from fetal to postnatal period, with a corresponding increase in production of organoids. Like organoids, spheroids show Wnt-dependent indefinite self-renewing properties but display a poorly differentiated phenotype reminiscent of incompletely caudalized progenitors. The spheroid transcriptome is strikingly different from that of adult intestinal stem cells, with minimal overlap of Wnt target gene expression. The receptor LGR4, but not LGR5, is essential for their growth. Trop2/Tacstd2 and Cnx43/Gja1, two markers highly enriched in spheroids, are expressed throughout the embryonic-day-14 intestinal epithelium. Comparison of in utero and neonatal lineage tracing using Cnx43-CreER and Lgr5-CreERT2 mice identified spheroidgenerating cells as developmental progenitors involved in generation of the prenatal intestinal epithelium. Ex vivo, spheroid cells have the potential to differentiate into organoids, qualifying as a fetal type of intestinal stem cell.