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Human gastric multi-regional assembloids for functional parietal maturation

ABSTRACT: Human gastric multi-regional assembloids for functional parietal maturation

PROVIDER: PRJNA1036239 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26686219_1.fastq.gz	Fastqsanger.gz
	SRR26686219_2.fastq.gz	Fastqsanger.gz
	SRR26686220_1.fastq.gz	Fastqsanger.gz
	SRR26686220_2.fastq.gz	Fastqsanger.gz
	SRR26686221_1.fastq.gz	Fastqsanger.gz

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Human gastric multi-regional assembloids for functional parietal maturation

Project description:Patient-derived human organoids have the remarkable capacity to self-organise into more complex structures. However, to what extent the gastric organoids can recapitulate the human stomach functions remains unexplored. Here, we report how gastric region-specific organoids can self-assemble into complex multi-regional assembloids showing levels of differentiation unattainable with simpler models. The assembloid shows preserved fundus, body and antrum regionality and gastric-specific crosstalk pathways arise. Remarkably, the increased complexity and cross communication between the different gastric regions, allows for the arise of the elusive parietal cell type, responsible for the production of gastric acid, the essential function for food digestion. We showed functional response to drugs targeting ATPase H+/K+ and further maturation of the assembloid when transplanted in vivo. This advanced in vitro model, using patient-derived tissue-specific progenitors, successfully recapitulates the structural and functional characteristics of the human stomach, offering a promising strategy for studying developmental processes, tissue interactions, and disease mechanisms that were previously challenging to attain.

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Human Multi-Regional Gastric Assembloids Favour Epithelial Cell Differentiation and Functional Parietal Maturation [paediatric patients]

Project description:Patient-derived human organoids have the remarkable capacity to self-organise into more complex structures. However, to which extend the gastric organoids can recapitulate the human stomach functions remains unexplored. Here, we report how gastric region-specific organoids can self-assemble into complex multi-regional assembloids showing levels of differentiation unattainable with simpler models. The assembloid shows preserved fundus, body and antrum regionality and gastric-specific crosstalk pathways arise. Remarkably, the increased complexity and cross communication between the different gastric regions, allows for the arise of the elusive parietal cell type, responsible for the production of gastric acid, the essential function for food digestion. We showed functional response to drugs targeting ATPase H+/K+ and further maturation of the assembloid when transplanted in vivo. This advanced in vitro model, using patient-derived tissue-specific progenitors, successfully recapitulates the structural and functional characteristics of the human stomach, offering a promising strategy for studying developmental processes, tissue interactions, and disease mechanisms that were previously challenging to attain.

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Human Multi-Regional Gastric Assembloids Favour Epithelial Cell Differentiation and Functional Parietal Maturation [paediatric patients]

Project description:Human Multi-Regional Gastric Assembloids Favour Epithelial Cell Differentiation and Functional Parietal Maturation [paediatric patients]

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Transcription profiling of mouse gastric epithelial cell types: parietal, zymogenic, and pit

Project description:Continuous regeneration of digestive enzyme (zymogen) secreting chief cells is a normal aspect of stomach function that is disrupted in pre-cancerous lesions. Regulation of zymogenic cell (ZC) differentiation is poorly understood. Here we profile Parietal, Pit, and Zymogenic cells for comparison and study. Experiment Overall Design: Samples were obtained through Laser-capture microdisection of gastric epithelium. The three samples come from enrichments for Zymogenic Cells (ZC), Parietal Cells (PC), and Mucus Pit Cells (Pit). Experiment Overall Design: Individual parietal cells (visualized by DBA-positivity and autofluorescence) from well-oriented gastric units were dissected (PixCell II LCM apparatus, 7.5-Âµm spot diameter; CapSure HS LCM caps, Arcturus, Mountain View, CA) one-at-a-time from the pit zone and collected for GeneChips. Pit cells (E-cadherin-positive, DBA-negative) were then collected from the same gastric units. Only the pit cells in a 2-3-cell-thick region at the apex of the gastric unit â?? but not yet upon the gastric surface â?? were taken. ZCs (E-cadherin-positive, GSII/DBA-negative cells in the base zones) were collected from different slides in corpus gastric units after potentially contaminating basal parietal cells had first been dissected and discarded. 3000-5000 individual cells from each cell lineage were isolated from 4-5 individual mice, and RNA was purified by PicoPure kit (Arcturus). RNA integrity was verified, and RNAs for each lineage were pooled from multiple mice and multiple dissections to make 20 ng total RNA, which was then amplified, labeled, and fragmented (by the Arcturus RiboAmp HS kit followed by the RNA Amplification and Labeling Kit from Enzo Life Sciences). The resulting biotinylated cRNA probes were hybridized to Affymetrix (Santa Clara, CA) GeneChipÂ® Mouse Genome 430 2.0 microarrays.

2008-06-13 | E-GEOD-5018 | biostudies-arrayexpress

A profile of murine gastric epithelial cells: Parietal, Zymogenic, Pit

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Microarray analysis of regenerating parietal cells (PCs) in murine gastric epithelium recovering from Diphtheria toxin injury

Project description:Highly specialized, acid-secreting PCs are one of many epithelial lineages that line the gastric epithelium. Even though PCs are functionally and structurally distinct from other lineages in the stomach, the molecular mechanisms that govern their specification and maturation from the gastric stem cells are relatively unknown. We used microarray analysis to interrogate transcriptional changes that occur in regenerating PCs and to identify candidate regulators of PC fate specification and maturation.

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Project description:Highly specialized, acid-secreting PCs are one of many epithelial lineages that line the gastric epithelium. Even though PCs are functionally and structurally distinct from other lineages in the stomach, the molecular mechanisms that govern their specification and maturation from the gastric stem cells are relatively unknown. We used single cell RNA sequencing (scRNA-seq) to construct a transcriptional profile for regenerating PCs and identify candidate regulators of PC fate specification and maturation.

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Gene expression in gastric Mist1+ cells.

Project description:Mist1+ cells and parietal cells in mouse stomach were separatedly sorted, and RNAs were isolated. Mist1 (also known as Bhlha15) is expressed in gastric chief cells and gastric stem cells in mice. However, more specific genes for each population needs to be identified to better understand the precise biology in these cell populations. In order to address cell specific gene signature, we separately sorted Mist1+ gastric chief cells and Mist1+ gastric stem cells by FACS, and performed microarray analysis. Mist1+ gastric chief cells were sorted by using Mist1-CreERT; R26-TdTomato mouse stomach, immediately after tamoxifen administration. Mist1+ gastric stem cells were sorted by chief cell-ablated Mist1-CreERT; R26-TdTomato mouse stomach, combining with Lgr5-DTR mice. Lgr5-expressing chief cells were ablated by giving DT into these mice. As a control, acid-secreting gastric parietal cell samples were used. Mice were treated with or without Lgr5-DT ablation before sorting.

2019-06-25 | GSE133205 | GEO

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