ETV2 Overexpression Promotes Efficient Differentiation of Pluripotent Stem Cells to Endothelial Cells
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ABSTRACT:
PROVIDER: PRJNA1129856 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR29653100_1.fastq.gz | Fastqsanger.gz | |||
| SRR29653100_2.fastq.gz | Fastqsanger.gz | |||
| SRR29653101_1.fastq.gz | Fastqsanger.gz | |||
| SRR29653101_2.fastq.gz | Fastqsanger.gz | |||
| SRR29653102_1.fastq.gz | Fastqsanger.gz |
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Project description:Directed differentiation of endothelial cells (ECs) from human pluripotent stem cells (hPSCs) typically takes around fourteen days, requires optimization of parameters such as hPSC seeding density and medium supplement concentration or timing, and often necessitates cell sorting to achieve high purity ECs. Overexpression of cell type-specific transcription factors in hPSCs has been shown to lead to efficient differentiation of many cell types, including neurons, astrocytes and hepatocytes. ETV2 has been identified as an essential transcription factor for endothelial cell fate commitment during development. Here, we present a simple, well-characterized differentiation method to generate ECs by overexpressing ETV2 in an inducible manner (iETV2) in hPSC cell lines. The result is a two-stage differentiation strategy, in which ECs are differentiated in a basal medium during stage I and then expanded in endothelial medium during stage II. By optimizing hPSC seeding density and differentiation medium, we generated an EC population with more than 99% pure CD31+ CD144+ cells without any cell sorting in just five days. In vitro angiogenesis potential of the iETV2-ECs was confirmed in cord formation assays and iETV2-EC uptake of acetylated LDL and response to inflammatory cytokines was confirmed. We compared transcriptomics of iETV2-ECs to the ECs generated from hPSCs via directed differentiation and confirmed similarity of gene expression profiles of ECs generated by these two different approaches. Furthermore, we found that iETV2-ECs respond to previously identified Wnt signaling agonist CHIR99021 and TGFβ signaling inhibitor RepSox to acquire aspects of brain EC phenotype. These findings underscore that iETV2-ECs can serve as easily scalable endothelial source, powering a variety of research applications, potentially including in vitro blood-brain barrier modeling.
2025-03-24 | GSE271101 | GEO
Project description:Etv2 transgene was expressed from ROSA26 locus by removing floxed STOP cassette by Vav Cre transgene. KSL or Mac1+/Gr1+ cells were sorted from control or Vav-Etv2 bone marrow and compared for gene expression in duplicate. This study will reveal the effect of Etv2 transgene in adult hematopoietic cells. The effect of Etv2 overexpression in relevant mouse tissue will be important to understand its effect in comparison with in ES cells. Genes aberrantly regulated by Etv2 overexpression will help to understand the caveat when using Etv2 to induce endothelial and hematopoietic cells in vitro. Sample ID #1;KSL;Control #2;KSL;Control #3;KSL;Tg #4;KSL;Tg #5;Gr;Control #6;Gr;Control #7;Gr;Tg #8;Gr;Tg
2012-05-09 | E-GEOD-36731 | biostudies-arrayexpress
Project description:Etv2 transgene was expressed from ROSA26 locus by removing floxed STOP cassette by Vav Cre transgene. KSL or Mac1+/Gr1+ cells were sorted from control or Vav-Etv2 bone marrow and compared for gene expression in duplicate. This study will reveal the effect of Etv2 transgene in adult hematopoietic cells. The effect of Etv2 overexpression in relevant mouse tissue will be important to understand its effect in comparison with in ES cells. Genes aberrantly regulated by Etv2 overexpression will help to understand the caveat when using Etv2 to induce endothelial and hematopoietic cells in vitro.
2012-05-10 | GSE36731 | GEO
Project description:Etv2 transgene was expressed from ROSA26 locus by removing floxed STOP cassette by Tie-2 Cre transgene.VE-Cad+/CD45+ cells were sorted from control or tie-2-Etv2 E10.5 embryosYS(yolk sac) and compared for gene expression in duplicate. This study will reveal the effect of Etv2 transgene in E10.5 mouse embryo yolk sac. The effect of Etv2 overexpression in relevant mouse tissue will be important to understand its effect in comparison with in ES cells. Genes aberrantly regulated by Etv2 overexpression will help to understand the caveat when using Etv2 to induce endothelial and hematopoietic cells in vitro. Sample ID YS- ; (VECAD+CD45+;YS;Control) YS+12; (VECAD+CD45+;YS;Tg) YS+22 ; (VECAD+CD45+;YS;Tg)
2012-05-09 | E-GEOD-36788 | biostudies-arrayexpress
Project description:Etv2 transgene was expressed from ROSA26 locus by removing floxed STOP cassette by Tie-2 Cre transgene.VE-Cad+/CD45+ cells were sorted from control or tie-2-Etv2 E10.5 embryosYS(yolk sac) and compared for gene expression in duplicate. This study will reveal the effect of Etv2 transgene in E10.5 mouse embryo yolk sac. The effect of Etv2 overexpression in relevant mouse tissue will be important to understand its effect in comparison with in ES cells. Genes aberrantly regulated by Etv2 overexpression will help to understand the caveat when using Etv2 to induce endothelial and hematopoietic cells in vitro.
2012-05-10 | GSE36788 | GEO
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
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
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
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.
2011-08-31 | GSE31743 | GEO
Project description:Mechanisms of cell fate specification are central to developmental biology and regenerative medicine. ETV2 is a master regulator for the endothelial cell lineage specification. Here, we study mechanisms by which ETV2 overexpression in human iPSC-derived mesodermal progenitors efficiently specifies ECs.
2025-04-01 | GSE267565 | GEO