2025-11-07
ENA first public
ERC000011
ENA-CHECKLIST
National Center for Cardiavascular Research (CNIC)
INSDC center name
public
INSDC status
ERS25058859
SRA accession
E-MTAB-15242:Sample_pool7cre_minus
Submitter Id
11
week
age
ArrayExpress
broker name
aorta cell
cell type
not collected
collection date
house mouse
common name
Protocols: Aortas from Ldlr-/- mice reconstituted with Lmna-/-, Lmna+/+, Lmna-OE, or Lmnatg BM were thoroughly perfused with PBS to remove blood. The aortic arch and thoracic aorta were dissected, cleaned of perivascular fat and opened longitudinally. Aortas from 3-5 transplant recipients of the same genotype were pooled, with both male and female donors included in each pool to minimize potential sex-related bias. Aortas were digested at 37°C for 15 minutes in DMEM supplemented with collagenase A (6.25 mg/ml, Roche), dispase II (6.25 mg/ml, Roche), DNase I (62.5 μg/ml; Roche), and elastase (1.717 U/ml, Sigma). Cells were treated with 0.25% trypsin-EDTA (Gibco) for 5 minutes at 37°C, pelleted and resuspended in sorting buffer. Purified cells were sorted to filter cells according to size and complexity (FSC, SSC), and then select viable nucleated cells (Hoechst 33342+, TO-PRO-3−) for sorting, discarding dead cells and debris. Cells were recounted and their viability checked on a Countess III cell counter (Thermofisher). Cells were loaded into each port of a Chromium Next GEM Chip G (10x Genomics) for single cell isolation and cDNA reverse transcription and amplification Sequencing libraries were created using the Next GEM Single cell 3’Library preparation kit v3.1 (10x Genomics) and indexed using the Chromium i7 Multiplex kit (10x Genomics) following the manufacturer instructions
description
adult
developmental stage
LmnaTG
genetic modification
Ldlr-/-; LmnaTG
genotype
not collected
geographic location (country and/or sea)
Pool4
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
aorta
organism part
Mus musculus
scientific_name
mixed
sex
C57BL/6
strain
Sample_pool7cre_minus
title