Plasticity of epithelial cells derived from human normal and ADPKD kidneys in primary cultures
Plasticity of epithelial cells derived from human normal and ADPKD kidneys in primary cultures
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DOI：
10.1007/s00441-007-0521-4
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发表时间：
2008-02-01
影响因子：
3.6
通讯作者：
Turman, Martin A.
中科院分区：
文献类型：
--
作者：
Elberg, Gerard;Guruswamy, Suresh;Turman, Martin A.
常染色体显性多囊肾病（ADPKD）的特征是肾小管上皮细胞去分化和增殖引发囊肿形成。原代培养物中的肾小管上皮细胞（RTC，源自正常肾组织）表现出γ-谷氨酰转移酶和低分子量细胞角蛋白的均质表达，这两种标记物分别代表近端和远端肾上皮细胞。培养物中的 RTC 还异常表达去分化标记物波形蛋白和 PAX-2，这些蛋白质通常在 ADPKD 肾脏囊肿内衬的上皮细胞中表达，但在正常肾脏的肾小管细胞中不表达。相比之下，囊性上皮细胞（CEC，源自多囊肾的囊壁）的不同培养物显示出细胞角蛋白、γ-谷氨酰转移酶和PAX-2的不同表达，但波形蛋白的表达水平恒定。重要的是，当在三维凝胶基质中培养时，RTC 和 CEC 表现出通过形成肾小管和囊肿分别转化为其各自原始结构的能力，而 HK-2、LLC-PK1 和 MDCK 肾上皮细胞系形成细胞聚集体或囊肿。我们的研究表明，各种上皮细胞类型的标记表达在原代培养物中并不高度稳定。它们的调节在源自正常和 ADPKD 肾脏的细胞以及单层和三维培养的细胞中是不同的。这些结果表明上皮细胞在培养扩增过程中表现出混合的上皮/去分化/间充质表型的可塑性。然而，三维培养中的 RTC 和 CEC 形态发生上皮特性与体内相似。因此，该模型可用于研究导致肾小管发生和囊肿发生的机制。
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by cyst formation initiated by dedifferentiation and proliferation of renal tubular epithelial cells. Renal tubular epithelial cells (RTC, derived from normal kidney tissue) in primary cultures exhibit both homogeneous expression of gamma-glutamyl transferase and low molecular weight cytokeratin, two different markers for proximal and distal renal epithelial cells, respectively. RTC in cultures also abnormally express the dedifferentiation markers vimentin and PAX-2, which are proteins normally expressed in epithelial cells lining cysts in ADPKD kidneys but not tubular cells in normal kidneys. In contrast, different cultures of cystic epithelial cells (CEC, derived from the cysts walls of polycystic kidneys) display variable expression of cytokeratin, gamma-glutamyl transferase, and PAX-2, but a constant level of vimentin. Importantly, RTC and CEC exhibit the capacity to convert to their respective original structures by forming tubules and cysts, respectively, when cultured in a three-dimensional gel matrix, whereas HK-2, LLC-PK1, and MDCK renal epithelial cell lines form cell aggregates or cysts. Our study demonstrates that the marker expression of the various epithelial cell types is not highly stable in primary cultures. Their modulation is different in cells originating from normal and ADPKD kidneys and in cells cultured in monolayer and three-dimensions. These results indicate the plasticity of epithelial cells that display a mixed epithelial/dedifferentiated/mesenchymal phenotype during their expansion in culture. However, RTC and CEC morphogenic epithelial properties in three-dimensional cultures are similar to those in vivo. Thus, this model is useful for studying the mechanisms leading to tubulogenesis and cystogenesis.