Identification of multipotent progenitors in the embryonic mouse kidney by a novel colony-forming assay
Identification of multipotent progenitors in the embryonic mouse kidney by a novel colony-forming assay
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DOI：
10.1242/dev.02174
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发表时间：
2006-01-01
期刊：
影响因子：
4.6
通讯作者：
Nishinakamura, R
中科院分区：
文献类型：
--
作者：
Osafune, K;Takasato, M;Nishinakamura, R
具有多向分化潜能的肾干细胞或祖细胞仍有待分离，这些细胞类型在肾脏发育或再生过程中的分化机制尚不清楚。为了解决这个问题，我们建立了一个体外培养系统，使用稳定表达Wnt 4的NIH 3 T3细胞（3 T3 Wnt 4）作为饲养层，其中后肾间充质中的单个肾祖细胞形成由存在于肾小球和肾小管中的几种类型的上皮细胞组成的集落。我们发现，只有细胞强烈表达Sall 1（Sall 1-GFp（高）细胞），锌指核因子对肾脏发育至关重要，形成集落，他们重建了一个三维肾脏结构在器官培养设置。我们还发现，Rac和JNK依赖的平面细胞极性（PCP）途径下游的Wnt 4正调控的殖民地的大小，JNK途径也参与间充质上皮转化的集落形成祖细胞。因此，我们的集落形成试验，它确定多能祖细胞在胚胎小鼠肾脏，可用于检查肾祖细胞分化的机制。
Renal stem or progenitor cells with a multilineage differentiation potential remain to be isolated, and the differentiation mechanism of these cell types in kidney development or regeneration processes is unknown. In an attempt to resolve this issue, we set up an in vitro culture system using NIH3T3 cells stably expressing Wnt4 (3T3Wnt4) as a feeder layer, in which a single renal progenitor in the metanephric mesenchyme forms colonies consisting of several types of epithelial cells that exist in glomeruli and renal tubules. We found that only cells strongly expressing Sall1 (Sall1-GFp(high) cells), a zinc-finger nuclear factor essential for kidney development, form colonies, and that they reconstitute a three-dimensional kidney structure in an organ culture setting. We also found that Rac- and JNK-dependent planar cell polarity (PCP) pathways downstream of Wnt4 positively regulate the colony size, and that the JNK pathway is also involved in mesenchymal-to-epithelial transformation of colony-forming progenitors. Thus our colony-forming assay, which identifies multipotent progenitors in the embryonic mouse kidney, can be used for examining mechanisms of renal progenitor differentiation.