U.S. flag

An official website of the United States government

Format

Send to:

Choose Destination

ERX3407644: Illumina HiSeq 4000 sequencing; Endothelial Cell Atlas
8 ILLUMINA (Illumina HiSeq 4000) runs: 237.6M spots, 23G bases, 10Gb downloads

Design: Endothelial Cell Atlas
Submitted by: Laboratory of Angiogenesis and Vascular Metabolism VIB-KU Leuven Center for Cancer Biology (CCB) (Laboratory of Angiogenesis and Vascular Metabolism)
Study: Endothelial Cell Atlas
show Abstracthide Abstract
The heterogeneity of endothelial cells (ECs), lining blood vessels, across tissues remains incompletely inventoried. We constructed an atlas of >32,000 single-EC transcriptomic data from 11 tissues of the model organism Mus musculus. We propose a new classification of EC phenotypes based on transcriptome signatures and inferred putative biological features. We identified top-ranking markers for ECs from each tissue. ECs from different vascular beds (arteries, capillaries, veins, lymphatics) resembled each other across tissues, but only arterial, venous and lymphatic (not capillary) ECs shared markers, illustrating a greater heterogeneity of capillary ECs. We identified high-endothelial-venule and lacteal-like ECs in the intestines, and angiogenic ECs in healthy tissues. Metabolic transcriptomes of ECs differed amongst spleen, lung, liver, brain and testis, while being similar for kidney, heart, muscle and intestines. Within tissues, metabolic gene expression was heterogeneous amongst ECs from different vascular beds, altogether highlighting large EC heterogeneity.
Sample: Small Intestine
SAMEA5723230 • ERS3526848 • All experiments • All runs
Organism: Mus musculus
Library:
Name: Small Intestine_p
Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC SINGLE CELL
Selection: PolyA
Layout: SINGLE
Construction protocol: Whole organs were dissected, followed by dissociation using tissue-specific digestion protocols. Samples were then enriched for endothelial cells (ECs) by magnetic bead sorting using the MACS system (Miltenyi Biotec). Next, positive selection of ECs was performed using CD31 MicroBeads (Miltenyi Biotec), followed by staining with various antibody combinations for multi-parameter flow acquisition. FACS-purification of ECs was then performed, and single cell suspensions of freshly isolated ECs were then resuspended in PBS containing 0.04% ultra-pure BSA. scRNA-seq libraries were prepared using the Chromium Single Cell 3′ Reagent Kits v2 (10x Genomics; Pleasanton, CA, USA) according to the manufacturer's instructions. The aimed target cell recovery for each library was 5000 (except for soleus, aimed recovery of 3000 cells). Generated libraries were sequenced on an Illumina HiSeq4000, followed by de-multiplexing and mapping to the mouse genome (build mm10) using CellRanger (10x Genomics, version 2.1.1). Samples were processed using V2 barcoding chemistry kits (10x Genomics). Single samples were always processed in a single well of a PCR plate, allowing all cells from a sample to be treated similarly. All samples were processed in parallel in the same thermal cycler.
Experiment attributes:
Experimental Factor: organism part: small intestine
Runs: 8 runs, 237.6M spots, 23G bases, 10Gb
Run# of Spots# of BasesSizePublished
ERR338356030,747,6393G1.3Gb2020-02-25
ERR338356127,453,5252.7G1.1Gb2020-02-25
ERR338356232,506,6233.2G1.3Gb2020-02-25
ERR338356327,177,2702.6G1.2Gb2020-02-25
ERR338360431,235,2833G1.3Gb2020-02-25
ERR338360527,954,5652.7G1.2Gb2020-02-25
ERR338360632,957,6593.2G1.3Gb2020-02-25
ERR338360727,589,5512.7G1.2Gb2020-02-25

ID:
10177837

Supplemental Content

Search details

See more...

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...