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A streamlined method to generate endothelial cells from human pluripotent stem cells via transient doxycycline-inducible ETV2 activation

ABSTRACT: A streamlined method to generate endothelial cells from human pluripotent stem cells via transient doxycycline-inducible ETV2 activation

PROVIDER: PRJNA1095525 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28536244_1.fastq.gz	Fastqsanger.gz
	SRR28536244_2.fastq.gz	Fastqsanger.gz
	SRR28536245_1.fastq.gz	Fastqsanger.gz
	SRR28536245_2.fastq.gz	Fastqsanger.gz
	SRR28536246_1.fastq.gz	Fastqsanger.gz

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Similar Datasets

A streamlined method to generate endothelial cells from human pluripotent stem cells via transient doxycycline-inducible ETV2 activation

Project description:The development of reliable methods for producing functional endothelial cells (ECs) is crucial for progress in vascular biology and regenerative medicine. In this study, we present a streamlined and efficient methodology for the differentiation of human induced pluripotent stem cells (iPSCs) into induced ECs (iECs) that maintain the ability to undergo vasculogenesis in vitro and in vivo using a doxycycline-inducible system for the transient expression of the ETV2 transcription factor. This approach mitigates the limitations of direct transfection methods, such as mRNA-mediated differentiation, by simplifying the protocol and enhancing reproducibility across different stem cell lines. We detail the generation of iPSCs engineered for doxycycline-induced ETV2 expression and their subsequent differentiation into iECs, achieving over 90% efficiency within four days. Through both in vitro and in vivo assays, the functionality and phenotypic stability of the derived iECs were rigorously validated. Notably, these cells exhibit key endothelial markers and capabilities, including the formation of vascular networks in a microphysiological platform in vitro and in a subcutaneous mouse model. Furthermore, our results reveal a close transcriptional and proteomic alignment between the iECs generated via our method and primary ECs, confirming the biological relevance of the differentiated cells. The high efficiency and effectiveness of our induction methodology pave the way for broader application and accessibility of iPSC-derived ECs in scientific research, offering a valuable tool for investigating endothelial biology and for the development of EC-based therapies.

2024-07-05 | GSE263058 | GEO

A streamlined method to generate endothelial cells from human pluripotent stem cells via transient doxycycline-inducible ETV2 activation.

Project description: Not available

| S-EPMC11577265 | biostudies-literature

RNA sequencing of hiPSC-derived endothelial cells via doxycycline-induced ETV2 transcription factor overexpression

Project description:Endothelial cells are known to express tissue-specific phenotypes to support the organ functionality due to the residing tissue microenvironment niche. The objective of this study is to compare the transcriptomic profiling of the generated endothelial cells (called iETV2-ECs) in neuro maintenance medium (NMM) with endothelial cells derived from primary human brain and other organs, as well as human induced pluripotent stem cells. The iETV2-ECs were generated from human induced pluripotent stem cells (hiPSC) that were genome engineered with a doxycycline-inducible overexpression cassette for the master regulator of endothelial differentiation (i.e. ETV2). The cells were treated with doxycycline for 8 days in the NMM supplemented with bFGF, FBS and endothelial cell growth supplements and subsequently maintained in NMM up to 21 days. Cells were harvested on day 8, 14 and 21 days for total RNA extraction, and their transcriptomic profiles were analyzed by bulk RNA sequencing technique. P ublicly-available RNA sequencing datasets of different endothelial cell types were integrated into the iETV2-EC data set for differentially expressed genes analysis. The ultimate goal of this project is to develop vascularized 3D neural model to study human brain vascularization and disease modelling.

2022-01-06 | GSE192919 | GEO

Comparison of Flk-1+/Etv2- vs Flk-1+/Etv2+ populations

Project description:Screening for genes up in Etv2+ cells within Flk-1+ ES derived mesoderm Microarray analysis performed to screen for the candidate genes regulated by Etv2. Differentiated Flk-1+ mesoderm can be devided into Etv2+ or-. Etv2+ cells are assumed to be committed to hemato/endothelial cells. Comparison of two populations can reveal genes relevant in this commitment. Extract RNA from sorted Flk-1+/Etv2- vs Flk-1+/Etv2+ populations.Etv2-Venus KI ES cells were differentiated on OP9 for 4-5 days and Flk-1+ population was separated into Etv2-Venus+ or- cells. Total RNA was purified from each population for analysis.

2011-08-30 | E-GEOD-31743 | biostudies-arrayexpress

Gene expression analysis of inducible ES cells overexpressing Etv2 (induced for 12 hours at day 3 of differentiation)

Project description:During embryogenesis, the endothelial and the hematopoietic lineages first appear during gastrulation in the blood island of the yolk sac. We have previously reported that an Ets variant gene 2 (Etv2/ER71) mutant embryo lacks hematopoietic and endothelial lineages, however, the precise roles of Etv2 in yolk sac development remains unclear. We carried out a transcriptome analysis using an ES cell line that can inducibly express Etv2. Cells were induced for 12 hours at day 3 of differentiation. An ES cell line (dervied from mouse of background strain 129P2/Ola) with a doxycycline inducible expression casette of Etv2 (ER71 A2Lox.cre) was differentiated in hanging drops. At 3 days of differentiation, cells were treated with doxycycline for 12 hours, and subsequently Flk1(+) cells were sorted. Experiment was done in triplicate and untreated cells were used for control.

2012-07-03 | E-GEOD-37658 | biostudies-arrayexpress

RNA sequencing of hiPSC-derived endothelial cells via doxycycline-induced ETV2 transcription factor overexpression

Project description:RNA sequencing of hiPSC-derived endothelial cells via doxycycline-induced ETV2 transcription factor overexpression

| PRJNA793821 | ENA

Comparison of Flk-1+/Etv2- vs Flk-1+/Etv2+ populations

2011-08-31 | GSE31743 | GEO

ETV2/ER71 regulates hematovascular lineage generation and vascularization through an H3K9 demethylase, KDM4A

Project description:ETV2/ER71, an ETS transcription factor, is critical for hematopoiesis and vascular development. However, knowledge on the molecular mechanisms behind ETV2-mediated gene transcription is limited. Here, we show that ETV2 together with KDM4A, an H3K9 demethylase, regulates hematopoietic and endothelial genes. Etv2-/- mouse embryonic stem cells (mESCs), which fail to generate hematopoietic and endothelial cells, showed enhanced levels of H3K9me3 on hematopoietic and endothelial genes. ETV2 interacts with KDM4A and the ETV2-mediated transcriptional activation of hematopoietic and endothelial genes is dependent on KDM4A histone demethylase activity. ETV2 and KDM4A co-occupy the transcription regulatory regions of genes whose expression is directly regulated by ETV2. Mice lacking Kdm4a and Etv2 in endothelial cells (Cdh5Cre;Kdm4af/f;Etv2f/f) displayed a more severe defect in perfusion recovery and neovascularization compared with Cdh5Cre;Kdm4af/f, Cdh5Cre;Etv2f/f mice and controls. Collectively, we demonstrated that ETV2 interacts with KDM4A and that this interaction is critical for FLK1+ cell generation, differentiation into the downstream lineages, and vascular regeneration.

2024-12-10 | GSE251753 | GEO

Single-cell RNA-seq analysis of wildtype and ETV2-overexpressing embryoid body-derived progenitor cells

Project description:Wild-type (WT) and Etv2-overexpressing (OE) mouse embryonic stem cells (mESC) were cultured and differentated to embryoid bodies (EBs) in mesodermal conditions. Induction of Etv2 in OE EBs was achieved by the addition of doxycycline. Cells were collected at different time points (i.e., D2, D3, D4 for WT and D3, D4 for OE EBs) following FACS based on the presence or absence of two surface markers: Flk1 (F+/-) and Pdgfrα (P+/-). We performed single-cell RNA sequencing (scRNA-seq) on the FACS sorted cells at different time points. The transcriptomic profiles revealed that ETV2 overexpression promotes HE lineage specification while repressing cardiac and skeletal muscle programs.

2025-06-03 | GSE298879 | GEO

ATAC-seq analysis of wildtype and ETV2-overexpressing embryoid body-derived progenitor cells

Project description:Wild-type (WT) and Etv2-overexpressing (OE) mouse embryonic stem cells (mESC) were cultured and differentated to embryoid bodies (EBs) in mesodermal conditions. Induction of Etv2 in OE EBs was achieved by the addition of doxycycline. Cells were collected at different time points (i.e., D2, D3, D4 for WT and D3, D4 for OE EBs) following FACS sorting based on the presence or absence of two surface markers: Flk1 (F+/-) and Pdgfrα (P+/-). ATAC-seq was performed using 50,000 cells and the Tn5 transposase enzyme. Sequencing of libraries was carried out on the Illumina NextSeq platform (2×50 bp). Chromatin accessibility profiling revealed that Etv2 overexpression during EB differentiation increases accessibility of genes related to the hematoendothelial lineage, while reducing accessibility of genes associated with the cardiac lineage.

2025-06-16 | GSE299629 | GEO

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