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Single-cell transcriptome of early hematopoiesis guides arterial endothelial-enhanced functional T cell generation from human PSCs

ABSTRACT: Single-cell transcriptome of early hematopoiesis guides arterial endothelial-enhanced functional T cell generation from human PSCs

PROVIDER: PRJNA608525 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11175547_1.fastq.gz	Fastqsanger.gz
	SRR11175547_2.fastq.gz	Fastqsanger.gz
	SRR11175548_1.fastq.gz	Fastqsanger.gz
	SRR11175548_2.fastq.gz	Fastqsanger.gz
	SRR11175549_1.fastq.gz	Fastqsanger.gz

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Single-cell transcriptome of early hematopoiesis guides arterial endothelial-enhanced functional T cell generation from human PSCs

Project description:Human pluripotent stem cells (hPSCs) provide a powerful platform for studying the dynamic molecular network towards hematopoiesis. To date, a comprehensive roadmap at single-cell resolution for hPSC-derived hematopoietic differentiation has not been established. Here, we performed extensive single-cell transcriptomic analyses to map fate choices and gene expression programs during hematopoietic differentiation of hPSCs and identified strategies to improve the quantity and quality of hPSC-derived hematopoietic cells. By focusing specifically on cell populations and molecular events involved in endothelial-to-hematopoietic transition (EHT), the EHT was identified accompanying by a decline in aerobic metabolism and providing hypoxia in vitro enhances arterial and hematopoietic differentiation. Furthermore, arterial endothelial cells are validated as a critical regulator of definitive hematopoietic progenitor specification. Collectively, our study provides benchmark datasets to understand the origins of human hematopoiesis and presents an advance for guiding the generation of functional hematopietic cells in vitro for clinical applications.

2021-09-22 | GSE145859 | GEO

Single-cell transcriptome of early hematopoiesis guides arterial endothelial-enhanced functional T cell generation from human PSCs.

Project description:[Figure: see text].

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Enhancement of Arterial Specification in Human Pluripotent Stem Cell Cultures Promotes Definitive Hematoendothelial Program with Broad Myelolymphoid Potential

Project description:Identification of the regulators that lead to arterial specification with definitive hematopoietic potential should help to design strategies to recapitulate HSC development from human pluripotent stem cells (hPSCs). Here, using ETS1 conditional H1 hESC line, we found that ETS1 induction at the mesodermal stage of differentiation dramatically enhances the arterial specification in hPSC cultures and formation of DLL4+CXCR4+/- arterial HE with lymphoid potential and the capacity to produce red blood cells with high expression of BCL11a and b-globin. ETS1 effect was mediated through upregulation of NOTCH signaling. Together, these findings demonstrated that promotion of arterial specification in cultures could aid in generation of definitive hematopoiesis from hPSCs.

2018-03-09 | GSE96815 | GEO

Human arterial endothelial cells from pluripotent stem cells for in vitro hematopoiesis and lymphopoiesis

Project description:Human arterial endothelial cells from pluripotent stem cells for in vitro hematopoiesis and lymphopoiesis

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Separated Pancreatic stellate cells (PSCs) by functional separation using modified Matrigel invasion assay

Project description:We separated PSCs by functional separation using moidfied Matrigel invasion assay our previously reported (Fujiwara, PLoS One, 2012). In this culture system, we evaluated two primary cultures of human PSCs. PSCs in the upper chambers were incubated for 72 hours in culture with SUIT-2 pancreatic cancer cells in the lower chambers, or as controls without pancreatic cancer cells in the lower chambers, and were collected from the top and bottom sides of the upper chambers separately. The top side-derived PSCs were cells that did not migrate toward the cancer cells, and the bottom side-derived PSCs were cells that migrated toward the pancreatic cancer cells. We then performed a DNA microarray analysis using these cells. The quality of the RNA samples was evaluated using Experion™ Automated Electrophoresis Station (Bio-Rad Laboratories, Hercules, CA). We used a HumanHT-12v4 Expression BeadChip (Illumina, San Diego, CA) for these analyses. The data were analyzed using Bead Studio software version 3.2.3 (Illumina).

2014-02-25 | E-GEOD-55300 | biostudies-arrayexpress

Leukemic hematopoiesis

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Gene expression of rat PSCs cultured on plastic, matrigel and collagen

Project description:Activated pancreatic stellate cells produce the fibrotic matrix in chronic pancreatitis and pancreatic cancer. In vitro protocols examining PSC biology have usually involved PSCs cultured on plastic, a non-physiological surface. However, PSCs cultured on physiological matrices e.g. MatrigelTM (normal basement membrane) and collagen (fibrotic pancreas), may have distinctly different behaviours compared to cells cultured on plastic. Therefore, we aimed to compare PSC gene expression after culture on plastic, MatrigelTM and collagen I. Total RNA from stellate cells in 10 cm Petri dishes was isolated by Qiagen RNeasy Mini Plus kit as per manufacturer’s instructions. The Agilent 2100 Bioanalyzer (Agilent Technologies Inc. Santa Clara, CA) was used for quality control of the isolated total RNA. Gene expression profiles of rat PSCs cultured on MatrigelTM, collagen I and plastic were analysed by whole rat genome microarray purchased from Affymetrix (Rat Gene 1.0 ST Array). This array was able to detect 27,342 rat genes, with approximately 26 probes on average per gene (referred to as a probe set).

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2011-09-29 | E-GEOD-28008 | biostudies-arrayexpress

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Project description:The presence of activated pancreatic stellate cells (PSCs) in the pancreatic ductal adenocarcinoma (PDAC) microenvironment plays a significant role in cancer progression. Macrophage migration inhibitory factor (MIF) is overexpressed in PDAC tissues and expressed by both cancer and stromal cells. The expression status of MIF and its receptors in PDAC- associated fibroblasts or PSCs and its pathophysiological roles are yet to be elucidated. The next-generation sequencing technique was adapted to check the effect of MIF absence on the expression of other genes in mice (mPSCs).

2022-09-28 | E-MTAB-9392 | biostudies-arrayexpress

Evaluating the transcriptional regulators of arterial gene expression via a catalogue of characterized arterial enhancers

Project description:The establishment and growth of the arterial endothelium requires the coordinated expression of numerous genes. However, regulation of this process is not yet fully understood. Here, we combined in silico analysis with transgenic models to characterize arterial-specific enhancers associated with eight key arterial identity genes (Acvrl1/Alk1, Cxcr4, Cxcl12, Efnb2, Gja4/Cx37, Gja5/Cx40, Nrp1 and Unc5b). Next, to elucidate the regulatory pathways upstream of arterial gene transcription, we investigated the transcription factors binding each arterial enhancer compared to a similar assessment of non-arterial endothelial enhancers. These results found that binding of SOXF and ETS factors was a common occurrence at both arterial and pan-endothelial enhancers, suggesting neither are sufficient to direct arterial specificity. Conversely, FOX motifs independent of ETS motifs were over-represented at arterial enhancers. Further, MEF2 and RBPJ binding was enriched but not ubiquitous at arterial enhancers, potentially marking an arterial sub-set. Lastly, there was no shared or arterial-specific signature for WNT-associated TCF/LEF, TGFβ/BMP-associated SMAD1/5 and SMAD2/3, shear stress-associated KLF4 or venous-enriched NR2F2. This cohort of well characterized and in vivo-verified enhancers can now provide a platform for future studies into the interaction of different transcriptional and signalling pathways with arterial gene expression.

2024-12-07 | GSE283369 | GEO

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