Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
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DOI：
10.1371/journal.pone.0034907
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Moeller MJ
中科院分区：
文献类型：
--
作者：
Kabgani N;Grigoleit T;Schulte K;Sechi A;Sauer-Lehnen S;Tag C;Boor P;Kuppe C;Warsow G;Schordan S;Mostertz J;Chilukoti RK;Homuth G;Endlich N;Tacke F;Weiskirchen R;Fuellen G;Endlich K;Floege J;Smeets B;Moeller MJ
壁上皮细胞（佩奇）参与了急进性肾小球肾炎（RPGN）以及局灶性节段性肾小球硬化（FSGS）的发病机制。在本研究中，使用转基因小鼠品系，使用FACsorting分离纯的、遗传标记的佩奇或足细胞原代培养物。通过这种方法，可以解析培养中的原代肾小球上皮细胞的形态：原代足细胞形成具有胞质内延伸的大细胞或较小的纺锤形细胞，这取决于特定的培养条件。原代佩奇较小，呈纺锤形或多边形形态。在原代培养的非常早期阶段，观察到基因表达（例如WT-1和Pax-2）的快速变化。然而，在长时间培养后，原代佩奇和足细胞在转录组分析中仍然明显分离-表明原代细胞培养物的来源是重要的。在经典的标记物中，突触足蛋白和podoplanin的表达在原代PEC和足细胞培养物中受到最大程度的差异调节。然而，没有任何内源性基因的表达允许在培养物中区分两种细胞类型。最后，我们表明，转录因子WT 1也表达的佩奇。总之，佩奇和足细胞的遗传标记是获得具有已证实来源的纯原代培养物的新颖且必要的工具。这些文化将是一个强大的工具，为新兴领域的壁上皮细胞生物学。
Parietal epithelial cells (PECs) are crucially involved in the pathogenesis of rapidly progressive glomerulonephritis (RPGN) as well as in focal and segmental glomerulosclerosis (FSGS). In this study, transgenic mouse lines were used to isolate pure, genetically tagged primary cultures of PECs or podocytes using FACsorting. By this approach, the morphology of primary glomerular epithelial cells in culture could be resolved: Primary podocytes formed either large cells with intracytoplasmatic extensions or smaller spindle shaped cells, depending on specific culture conditions. Primary PECs were small and exhibited a spindle-shaped or polygonal morphology. In the very early phases of primary culture, rapid changes in gene expression (e.g. of WT-1 and Pax-2) were observed. However, after prolonged culture primary PECs and podocytes still segregated clearly in a transcriptome analysis - demonstrating that the origin of primary cell cultures is important. Of the classical markers, synaptopodin and podoplanin expression were differentially regulated the most in primary PEC and podocyte cultures. However, no expression of any endogenous gene allowed to differentiate between the two cell types in culture. Finally, we show that the transcription factor WT1 is also expressed by PECs. In summary, genetic tagging of PECs and podocytes is a novel and necessary tool to derive pure primary cultures with proven origin. These cultures will be a powerful tool for the emerging field of parietal epithelial cell biology.
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影响因子：
19.6
作者：
Le Hir, M;Besse-Eschmann, V
通讯作者：
Besse-Eschmann, V
影响因子：
19.6
作者：
KRIZ, W;HAHNEL, B;ELGER, H
通讯作者：
ELGER, H
DOI：
10.1097/01.asn.0000102468.37809.c6
发表时间：
2004-01-01
影响因子：
13.6
作者：
Moeller, MJ;Soofi, A;Holzman, LB
通讯作者：
Holzman, LB
影响因子：
19.6
作者：
Dijkman, H;Smeets, B;Wetzels, J
通讯作者：
Wetzels, J
影响因子：
13.6
作者：
Ronconi, Elisa;Sagrinati, Costanza;Romagnani, Paola
通讯作者：
Romagnani, Paola