2024-08-08
ENA first public
ERC000011
ENA-CHECKLIST
Centro Nacional de Investigaciones Cardiovasculares Carlos III (F.S.P.)
INSDC center name
public
INSDC status
ERS20599133
SRA accession
E-MTAB-14252:rHDL
Submitter Id
16
week
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Aortic arch and root samples from Apoe-/- mice fed a Western diet for 6 weeks were collected after treatment with either rHDL or PS@rHDL (n = 6 per group). Mice were fasted for 24 hours and following euthanasia were perfused with 5 mM EDTA PBS (Sigma-Aldrich, E1644). After adipose tissue removal, aortic arch and root samples were diced and digested with a cocktail of enzymes, including liberase TH (4 U/mL; Sigma-Aldrich, 5401151001), hyaluronidase type I-S (60 U/mL; Sigma-Aldrich, H3506) and deoxyribonuclease I (40 U/mL; Sigma-Aldrich, DN25) in PBS at 37ºC for 1 hour in an Eppendorf ThermoMixer C thermoblock shaking at 300 rpm. Cells were resuspended in 2% fetal bovine serum (FBS) PBS and passed through 70-μm cell strainers (VWR, 734-2761) to obtain single-cell suspensions, which were pooled into one sample per treatment group. Cells were then incubated with DAPI (Sigma-Aldrich, D8417) and DRAQ5 (Invitrogen, 65-0880-96) for dead cell exclusion. Cells were sorted using a FACS Aria II cell sorter equipped with BD FACSDivaTM Software v6.1.3. Cells were counted and their viability was checked using the Countess 3 cell counter (ThermoFisher). Mice were randomly allocated to two treatment groups, receiving rHDL (~20 mg apoA1/kg, i.v.) or PS@rHDL (20 mg PS/kg, i.v.). Doses were administered via lateral tail vein injection. The treatment schedule consisted of 8 administrations over two weeks, during which time mice were kept on Western diet. Animals were fasted from the time of last administration until euthanasia 24 hours later, following which they were perfused with PBS and tissues of interest collected. N/A. No nucleic extraction needed as this was a single-cell transcriptomics experiment. Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics).
description
adult
developmental stage
Ssniff E15721-347 EF TD88137
diet
Apoe-/-
genotype
not collected
geographic location (country and/or sea)
Pool1
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
Aortic arch and root
organism part
Mus musculus
scientific_name
female
sex
C57BL/6J
strain
rHDL
title