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A modular differentiation system maps multiple human kidney lineages from pluripotent stem cells [scRNA-Seq]

ABSTRACT: Recent studies using human pluripotent stem cells (hPSCs) have developed protocols to induce kidney-lineage cells and reconstruct kidney organoids. However, the separate generation of metanephric nephron progenitors (NPs), mesonephric NPs and ureteric bud (UB) cells, which constitute embryonic kidneys, in in vitro differentiation culture systems has not been fully investigated. Here we created a culture system in which these mesoderm-like cell types as well as paraxial and lateral plate mesoderm-like cells were separately generated from hPSCs. We succeeded in recapitulating nephrogenic niches from separately induced metanephric NP-like and UB-like cells, which were subsequently differentiated into glomeruli, renal tubules and collecting ducts in vitro and further vascularized in vivo. Our selective differentiation protocols should contribute to understanding the mechanisms underlying human kidney development and diseases and also supply cell sources for regenerative therapies.

ORGANISM(S): Homo sapiens

PROVIDER: GSE146118 | GEO | 2020/07/15

REPOSITORIES: GEO

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A modular differentiation system maps multiple human kidney lineages from pluripotent stem cells [RNA-Seq]

Project description:Recent studies using human pluripotent stem cells (hPSCs) have developed protocols to induce kidney-lineage cells and reconstruct kidney organoids. However, the separate generation of metanephric nephron progenitors (NPs), mesonephric NPs and ureteric bud (UB) cells, which constitute embryonic kidneys, in in vitro differentiation culture systems has not been fully investigated. Here we created a culture system in which these mesoderm-like cell types as well as paraxial and lateral plate mesoderm-like cells were separately generated from hPSCs. We succeeded in recapitulating nephrogenic niches from separately induced metanephric NP-like and UB-like cells, which were subsequently differentiated into glomeruli, renal tubules and collecting ducts in vitro and further vascularized in vivo. Our selective differentiation protocols should contribute to understanding the mechanisms underlying human kidney development and diseases and also supply cell sources for regenerative therapies.

2020-07-15 | GSE146117 | GEO

Generation of nephronal cells from human pluripotent stem cells via renal-vesicle formation

Project description:Human pluripotent stem cells (hPSC) provide a possible source for generation of kidney cells and organoids applicable in regenerative medicine, disease modeling and drug screening. By analyzing the effect of different factors on renal differentiation, we established an 8-day-protocol that steered hPSCs to the renal lineage by a step-wise process, segmented into mesoderm induction, intermediate mesoderm specification and metanephric induction outlining renal organogenesis. The differentiated cells contain populations of SIX2+/CITED1+ metanephric mesenchyme- (MM) and of HOXB7+/GRHL2+ ureteric bud (UB)-like cells at the end of 6 days. Transcriptome analysis corroborated that the in vitro generated precursor cell types at day 8 resemble their renal vesicle counterparts in vivo. The cells were further differentiated in 2-dimensional culture into podocyte- and diverse tubular epithelial-like cell types, showing their nephrogenic potential. In 3-dimensional culture, the progenitors gave rise to renal vesicles, and upon mouse embryonic kidney re-aggregation they form tubular structures.

2018-11-08 | GSE75711 | GEO

Expression data from mouse embryonic ureteric bud (UB) cells

Project description:Dicer-dependent miRNAs are required for UB morphogenesis and differentiation during metanephric kidney development. We used microarray analysis to identify genes whose expression in the UB epithelium are altered from UB-specific ablation of Dicer.

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Expression data from mouse embryonic ureteric bud (UB) cells

2014-08-27 | E-GEOD-60758 | biostudies-arrayexpress

Transcriptomically-guided mesendoderm induction of human pluripotent stem cells using a systematically defined culture scheme

Project description:Human pluripotent stem cells (hPSCs) are an essential cell source in tissue engineering, studies of development, and disease modeling. Efficient and broadly amenable protocols for rapid lineage induction of hPSCs are of great interest in the stem cell biology field. We describe a simple yet robust method for differentiation of hPSCs into mesendoderm in defined conditions utilizing single-cell seeding and BMP4 and Activin A (BA) treatment. Gene sets and gene ontology terms related to mesoderm and endoderm differentiation were enriched after 48 hours of BA treatment. After 14 days of differentiation, BA-treated cells expressed genes indicative of mesoderm and endoderm-derived tissue types, including lung, kidney and adipose tissue. BA treatment was readily incorporated into existing protocols for chondrogenic and endothelial progenitor cell differentiation. Teratomas formed from BA-treated cells contained higher ratios of mesoderm and endoderm to ectoderm tissue compared to PSC-derived teratomas. Collectively, these data demonstrate that single-cell seeding with BA treatment is a powerful method for induction of mesendoderm that can be integrated into protocols for mesoderm and endoderm differentiation.

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Direct Isolation and Characterization of Human Nephron Progenitors.

Project description:Mature nephrons originate from a small population of uninduced nephrogenic progenitor cells (NPs) within the cap mesenchyme. These cells are characterized by the coexpression of SIX2 and CITED1. Many studies on mouse models as well as on human pluripotent stem cells have advanced our knowledge of NPs, but very little is known about this population in humans, since it is exhausted before birth and strategies for its direct isolation are still limited. Here we report an efficient protocol for direct isolation of human NPs without genetic manipulation or stepwise induction procedures. With the use of RNA-labeling probes, we isolated SIX2+CITED1+ cells from human fetal kidney for the first time. We confirmed their nephrogenic state by gene profiling and evaluated their nephrogenic capabilities in giving rise to mature renal cells. We also evaluated the ability to culture these cells without complete loss of SIX2 and CITED1 expression over time. In addition to defining the gene profile of human NPs, this in vitro system facilitates studies of human renal development and provides a novel tool for renal regeneration and bioengineering purposes

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BET proteins regulate gene expression in early kidney development

Project description:In utero renal development is subject to maternal metabolic and environmental influences af-fecting long-term renal function and the risk to develop chronic kidney failure and cardiovascular disease. Epigenetic processes have been implicated in the orchestration of renal development and prenatal programming of nephron number. Bromodomain and extraterminal domain proteins act as histone acetylation reader molecules and promote gene transcription. BET family members Brd2, Brd3 and Brd4 are expressed in the nephrogenic zone during kidney development. Here, the effect of FDA-approved BET inhibitor JQ1 on renal development is evaluated. Inhibition of BET proteins via JQ1 leads to reduced growth of metanephric cultures, loss of the progenitor cell population, and premature and disturbed nephron differentiation.

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The developmental programme for genesis of the entire kidney is recapitulated in Wilms tumour

Project description:Expression profiling of Wilms tumour samples, which identified molecular signatures of the ureteric bud and collecting duct as well as those of the proximal and distal tubules in triphasic histology tumours. These observations indicate Wilms tumours can arise from a precursor cell capable of generating the entire kidney, such as the cells of the intermediate mesoderm from which both the metanephric mesenchym and ureteric bud are derived.

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10.5dpc Metanephric Mesenchyme V Intermediate Mesoderm

Project description:10.5dpc Metanephric Mesenchyme V Intermediate Mesoderm Keywords: repeat sample

2005-07-20 | GSE2301 | GEO

mRNA sequencing of wildtype versus CEP83 knockout hiPSCs at diffirent time points of differentiation to kidney organoids [scRNA-seq]

Project description:Centrosomal protein 83 (CEP83) is a component of the distal appendages proteins of centrioles, which is necessary for the assembly of primary cilia. Previous studies have implicated primary cilia in normal development and tissue homeostasis, including kidney development. The kidney develops from human induced pluripotent stem cell (hiPSCs) in a stepwise process involving induction of specialized Nephron progenitors (NPs) in the intermediate mesoderm (IM), followed by their differentiation into kidney epithelia. Here, we used CRISPR-cas9 technology to knockout CEP83 in hiPSCs that were differentiated versus the wildtype hiPSCs into IM followed by kidney epithelial differentiation to analyze the role of CEP83 in human kidney epithelial differentiation. We used morphological analyses, gene expression studies and immunostaining to analyze IM and nephron differentiation. Bulk RNA and single cell sequencing showed that CEP83-/- IM cells abnormally upregulate regulatory transcription factors of lateral plate mesoderm (LPM) including OSR1, FOXF1, FOXF2, HAND2 and FEDRR., and downregulate marker genes in specific NPs ncluding; PAX8, HOXB7 and EYA1. Further, wildtype hiPSCs successfully differentiated into kidney organoids that upregulate key nephron markers, including NPHS1, CUBN and GATA3. In contrast, CEP83-/- hiPSCs failed to differentiate into nephron epithelia and didn’t express nephron marker genes. In a human system modeling kidney epithelial differentiation from hiPSCs, our data provide a new insight to the essential role of CEP83 in NPs differentiation and may help to better understand the pathogenesis of CEP83-associated renal developmental defects.

2022-10-12 | GSE205977 | GEO

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