A semi-permeable insert culture model for the distal part of the nephron with human and mouse tubuloid epithelial cells

Tubuloids are advanced in vitro models obtained from adult human or mouse kidney cells with great potential for modelling kidney function in health and disease. Here, we developed a polarized human and mouse tubuloid epithelium on cell culture inserts, namely Transwell™ filters, as a model of the di...

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Bibliographic Details
Published in:	Experimental cell research Vol. 444; no. 1; p. 114342
Main Authors:	Dilmen, E., Olde Hanhof, C.J.A., Yousef Yengej, F.A., Ammerlaan, C.M.E., Rookmaaker, M.B., Orhon, I., Jansen, J., Verhaar, M.C., Hoenderop, J.G.
Format:	Journal Article
Language:	English
Published:	Elsevier Inc 01-01-2025
Subjects:	Epithelium Kidney Organoids Stem cell Tubuloids Organoids Tubuloids Epithelium Stem cell Kidney
Online Access:	Get full text
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Summary:	Tubuloids are advanced in vitro models obtained from adult human or mouse kidney cells with great potential for modelling kidney function in health and disease. Here, we developed a polarized human and mouse tubuloid epithelium on cell culture inserts, namely Transwell™ filters, as a model of the distal nephron with an accessible apical and basolateral side that allow for characterization of epithelial properties such as leak-tightness and epithelial resistance. Tubuloids formed a leak-tight and confluent epithelium on Transwells™ and the human tubuloids were differentiated towards the distal part of the nephron. Differentiation induced a significant upregulation of mRNA and protein expression of crucial segment transporters/channels NKCC2 (thick ascending limb of the loop of Henle), NCC (distal convoluted tubule), AQP2 (connecting tubule and collecting duct) and Na+/K+-ATPase (all segments) in a polarized fashion. In conclusion, this study illustrates the potential of human and mouse tubuloid epithelium on Transwells™ for studies of tubuloid epithelium formation and tubuloid differentiation towards the distal nephron. This approach holds great promise for assisting future research towards kidney (patho)physiology and transport function. •Kidney organoids and tubuloids are emerging cell models to study (patho)physiology.•Both mouse and human tubuloids can be cultured as monolayers on Transwell filters.•Mouse and human monolayers differentiate towards the distal part of the nephron.•Mouse and human tubuloid monolayers express polarized segment-specific markers.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0014-4827 1090-2422 1090-2422
DOI:	10.1016/j.yexcr.2024.114342