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Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential [DiI-VLDL]

ABSTRACT: Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential [DiI-VLDL]

PROVIDER: PRJNA639512 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12015805_1.fastq.gz	Fastqsanger.gz
	SRR12015805_2.fastq.gz	Fastqsanger.gz
	SRR12015806_1.fastq.gz	Fastqsanger.gz
	SRR12015806_2.fastq.gz	Fastqsanger.gz

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Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential [DiI-VLDL]

Project description:Ureteric bud (UB) is the embryonic kidney progenitor tissue that gives rise to the collecting duct and lower urinary tract. UB-like structures generated from human pluripotent stem cells by previously reported methods show limited developmental ability and limited branching. Here we report a new method to generate UB organoids that possess epithelial polarity and tubular lumen and repeat branching morphogenesis. We also succeeded in monitoring UB tip cells by utilizing the ability of tip cells to uptake very-low-density lipoprotein, cryopreserving UB progenitor cells and expanding UB tip cells that can reconstitute the organoids and differentiate into collecting duct progenitors. Moreover, we successfully reproduced some phenotypes of multicystic dysplastic kidney (MCDK) using the UB organoids. These methods will help elucidate the developmental mechanisms of UB branching and develop a selective differentiation method for collecting duct cells, contributing to the creation of disease models for congenital renal abnormalities.

2020-06-16 | GSE152471 | GEO

Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential

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Expansion of human iPSC derived ureteric bud organoids with repeated branching potential

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HNF1B controls epithelial organization and cell polarity during ureteric bud branching and collecting duct morphogenesis

Project description:Kidney development depends critically on proper ureteric bud branching giving rise to the entire collecting duct system. The transcription factor HNF1B is required for the early steps of ureteric bud branching. Yet, the molecular and cellular events regulated by HNF1B are poorly understood. We report that specific removal of Hnf1b from the ureteric bud leads to defective cell-cell contacts and apico-basal polarity during the early branching events. High resolution ex vivo imaging combined with a membranous fluorescent reporter strategy show decreased mutant cell-rearrangements during mitosis-associated cell dispersal and severe epithelial disorganisation. Molecular analysis reveals downregulation of Gdnf-Ret pathway components and suggests that HNF1B acts both upstream and downstream of Ret-signaling by directly regulating Gfrα1 and Etv5. Subsequently, Hnf1b-deletion leads to massively mispatterned ureteric tree network, defective collecting duct differentiation and disrupted tissue architecture leading to cystogenesis. Consistently, mRNA-seq analysis performed from E15.5 control and mutant kidneys shows that the most impacted genes encode intrinsic cell-membrane components with transporter activity. Our study uncovers a fundamental and recurrring role of HNF1B in epithelial organization during early ureteric bud branching and further patterning and differentiation of the collecting duct system. Article submitted to Development. Authors: Audrey Desgrange, Claire Heliot,Ilya Skovorodkin, Saad U. Akram, Janne Heikkilä, Veli-Pekka Ronkainen, Ilkka Miinalainen I., Seppo J. Vainio and Silvia Cereghini

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Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential [CDP & TIP]

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Generation of kidney ureteric bud and collecting duct organoid

Project description:A platform for generating expandable, branching and gene-editable ureteric bud organoid from primary mouse and human ureteric bud progenitor cells and human pluripotent stem cells, and its maturation into collecting duct organoid.

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Expansion of human iPSC derived ureteric bud organoids with repeated branching potential

Project description:We report a new method to generate UB organoids that possess epithelial polarity and tubular lumen and repeat branching morphogenesis. We also succeeded in monitoring UB tip cells by utilizing the ability of tip cells to uptake very-low-density lipoprotein, cryopreserving UB progenitor cells and expanding UB tip cells that can reconstitute the organoids and differentiate into collecting duct progenitors. Moreover, we successfully reproduced some phenotypes of multicystic dysplastic kidney (MCDK) using the UB organoids.

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Hepatocyte nuclear factor 1β gene regulation in E14.5 ureteric bud cells

Project description:HNF-1β mutations are one of the most common single-gene mutations that underlie kidney developmental disease. Hepatocyte nuclear factor 1β (HNF-1β) is essential for kidney development, but its functions in ureteric bud (UB) branching morphogenesis are incompletely understood. We isolated E14.5 UB cells using fluorescence-activated cell sorting, and performed RNA-sequencing to compare gene expression in wild-type and HNF-1β deficient UB cells. 1632 genes have significantly lower expression in the HNF-1β deficient UB cells and 2223 genes have significantly higher expression in HNF-1β deficient UB cells.

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