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Author

Kwon, H. Moo
Inflammation and Kidney Disorder Lab
Research Interests
  • TonEBP, Obesity, Cancer, Chronic inflammatory diseases, Brain disorder, Kidney disorders, Genomic instability

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Roles of fluid shear stress and retinoic acid in the differentiation of primary cultured human podocytes

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Title
Roles of fluid shear stress and retinoic acid in the differentiation of primary cultured human podocytes
Author
Yang, Seung HeeChoi, Jin WooHuh, DongeunJo, Hyung AhKim, SejoongLim, Chun SooLee, Jung ChanKim, Hee ChanKwon, H. MooJeong, Chang WookKwak, CheolJoo, Kwon WooKim, Yon SuKim, Dong Ki
Keywords
Podocyte;  Differentiation;  Microfluidics;  Retinoic acid
Issue Date
201705
Publisher
ELSEVIER INC
Citation
EXPERIMENTAL CELL RESEARCH, v.354, no.1, pp.48 - 56
Abstract
Due to the distinct features that distinguish immortalized podocyte cell lines from their in vivo counterparts, primary cultured human podocytes might be a superior cell model for glomerular disease studies. However, the podocyte de-differentiation that occurs in culture remains an unresolved problem. Here, we present a method to differentiate primary cultured podocytes using retinoic acid (RA) and fluid shear stress (FSS), which mimic the in vivo environment of the glomerulus. RA treatment induced changes in the cell shape of podocytes from a cobblestone-like morphology to an arborized configuration with enhanced mobility. Moreover, the expression of synaptopodin and zonula occludens (ZO)-1 in RA-treated podocytes increased along with Krüppel-like factor 15 (KLF15) expression. Confocal microscopy revealed that RA increased the expression of cytoplasmic synaptopodin, which adopted a filamentous arrangement, and junctional ZO-1 expression, which showed a zipper-like pattern. To elucidate the effect of FSS in addition to RA, the podocytes were cultured in microfluidic devices and assigned to the static, static+RA, FSS, and FSS+RA groups. The FSS+RA group showed increased synaptopodin and ZO-1 expression with prominent spikes on the cell-cell interface. Furthermore, interdigitating processes were only observed in the FSS+RA group. Consistent with these data, the mRNA expression levels of synaptopodin, podocin, WT-1 and ZO-1 were synergistically increased by FSS and RA treatment. Additionally, the heights of the cells were greater in the FSS and FSS+RA groups than in the static groups, suggesting a restoration of the 3D cellular shape. Meanwhile, the expression of KLF15 increased in the RA-treated cells regardless of fluidic condition. Taken together, FSS and RA may contribute through different but additive mechanisms to the differentiation of podocytes. These cells may serve as a useful tool for mechanistic studies and the application of regenerative medicine to the treatment of kidney diseases.
URI
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DOI
http://dx.doi.org/10.1016/j.yexcr.2017.03.026
ISSN
0014-4827
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