2025-07-09
ENA first public
ERC000011
ENA-CHECKLIST
Spanish National Center for Cardiovascular Research
INSDC center name
public
INSDC status
ERS18415079
SRA accession
E-MTAB-13898:Sample_Pig1_Aorta
Submitter Id
15
week
age
ArrayExpress
broker name
not collected
collection date
pig
common name
Protocols: Animals were euthanized and the abdominal aorta and iliac arteries were quickly isolated and opened. The atherosclerotic plaque was dissected from the distal 0.5 cm and proximal 0.5 cm of the right iliac artery. The media, adventitia, and perivascular tissue were manually removed and approximately 200-250 mg of plaque was selected, washed in HBSS, and minced. It was incubated for 60 minutes at 37 °C in 1 ml of an enzyme solution containing 1,5 mg/ml (8 U/ml) of elastase (LS002279, Worthington Biochemical), 2 mg/ml of liberase (05401119001, Sigma Aldrich) and 300 µg/ml of DNaseI (Sigma Aldrich, DN25). Non-digested tissue was disaggregated using a smooth-tipped Pasteur pipette. Reactions were stopped by adding PBS containing 0.5% bovine serum albumin (BSA). The suspension was passed through a 70 µm pore filter to remove aggregates. Cells were collected by centrifugation and  resuspended in 500 µl PBS+0.5% BSA. DAPI (final concentration, 1 µg/ml final ) and Draq5 (Thermo Scientific 65-0880-96; final concentration, 5 µM) were added to enable detection of viable and dead cells. Viable cells (Draq5 positive and DAPI negative) were sorted with a FACSAria Cell Sorter.  BMS-212122 is a chemical inhibitor for Microsomal Triglyceride Transfer Protein (MTP) with the IUAPC name 9- {4- [2,5-Dimethyl- 4- ({[4'-(trifluoromethyl)-2-biphenylyl] carbonyl} amino)-1H-benzimidazol-1-yl] butyl}-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide.Atherosclerosis was induced in PCSK9D374Y transgenic minipigs (males) by feeding a standard pig feed supplemented with 20% w/w lard, 1.5% w/w cholesterol (HFHC), 0.7% NaCl w/w, from 3 until 15 months of age. During the first 6 months, the diet was also supplemented with 0.7% cholic acid. Animals were then either continued on the HFHC diet for 3 months (HFHC group) or changed to the standard pig feed with supplemented BMS-212122 (LF/MTPi group). The BMS-212122 dose was 1 mg/(kg·day)  during the first week and 0.5 mg/(kg·day) for the rest of intervention period. Up to 16500  cells were loaded into each port of a Chromium Next GEM Chip G (10x Genomics). After reverse transcription and cDNA amplification, sequencing libraries were prepared using the Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics).
description
adult
developmental stage
high fat diet
diet
atherosclerosis
disease
PCSK9^D374Y
genotype
not collected
geographic location (country and/or sea)
HFHC
growth condition
not applicable
isolate
Sus scrofa
organism
Sus scrofa
organism
abdominal aorta
organism part
Sus scrofa
scientific_name
female
sex
Sample_Pig1_Aorta
title