Sample 2
Mus musculus
Protocols: Aortas were dissected and digested for 30 mins at 37°C in 1mg/ml collagenase I (Gibco, Life Technologies, NY, USA), 1mg/ml Dispase (Gibco, Life Technologies, NY, USA), 150μg/ml DNase-I (Sigma-Aldrich, St Louis, MO, USA) before passing the suspension through a 70μM cell strainer. Dissociated single cells in PBS/BSA/EDTA were then incubated with various antibody combinations for multi-parameter flow acquisition and analysis. The following combinations of antibodies were used to assess the endothelial hierarchy populations: Rat anti-mouse VEGFR2 PE, CD31 PE-Cy7 and CD34 Alexa647 (Becton Dickinson, NJ, USA), Rat anti-mouse Lineage cocktail BV450 (Biolegend), Rat anti-mouse CD144 FITC (eBioscience). FACS was performed on a FACSAria cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA). scRNA-seq was performed in triplicate using wild-type C57/BL6 mice. The 10X Genomics Chromium instrument (10X Genomics) was used to partition viable cells with barcoded beads, and cDNA from each cell was prepared using the Single Cell 3' Library, Gel Bead and Multiplex Kit (v2; 10X Genomics) as per the manufacturer's instructions. Library construction performed as per 10X Genomic Single Cell 3' RNA-seq protocol version 2
ENA-FIRST-PUBLIC
2019-03-31T17:01:55Z
ENA-LAST-UPDATE
2018-08-16T11:22:52Z
External Id
SAMEA4832938
INSDC center name
Division of Genomics of Development and Disease, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia, 4072
INSDC first public
2019-03-31T17:01:55Z
INSDC last update
2018-08-16T11:22:52Z
INSDC status
public
Submitter Id
E-MTAB-7149:Sample 2
age
6 to 8
broker name
ArrayExpress
cell type
aortic endothelial cell
common name
house mouse
disease
normal
genotype
wild type genotype
organism part
aorta
replicate
2
sample name
E-MTAB-7149:Sample 2
scientific_name
Mus musculus
sex
female
strain
C57BL/6