2025-05-01
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA113322251
External Id
Institut fur Pharmakologie Medizinische Fakultat der Heinrich-Heine-Universitat Dusseldorf
INSDC center name
2023-05-02T13:32:37Z
INSDC last update
public
INSDC status
ERS15317412
SRA accession
E-MTAB-12880:Rot5KO
Submitter Id
12
week
age
ArrayExpress
broker name
cardiac interstitial cell
cell_type
house mouse
common name
Protocols: Cardiac interstitial cells were isolated from WT and Vcan-KO mice (n=2,3) 7 days after cardiac ischemia with reperfusion. 10 min prior to harvest, mice were injected with 250 IU heparin i.p., hearts were perfused with cold PBS for 2 min before removal to a glass dissection plate. Atria and valves were discarded and ventricular tissue was moved to a fresh 100 mm petri dish on ice with a cold droplet of PBS. Ventricles were then minced until 1 mm cubes and then moved to a pre-chilled 2 mL tube containing a digestion solution, consisting of 600 U/mL collagenase type I (Worthington) and 60 U/mL DNase I (Sigma) in HBSS with calcium and magnesium. Tissue was then digested at 37 C for 20 min while rocking, with vigorous pipetting to disrupt the tissue every 10 min. The cell suspension was then filtered over a 100 µm filter sitting in a 50 mL falcon tube, the volume was then adjusted to 15 mL with cold PBS before filtering again with a 40 µm filter. Cells were then pelleted at 800 g for 10 min at 4 C. Supernatant was discarded, and erythrocytes were lysed for 7 min at 4 C. Cells were pelleted again at 300 g for 10 min at 4 C, resuspended in PEB, and incubated with 7-Aminoactinomycin D (Invitrogen), to excluded dead cells and anti-CD31 and anti-CD45 antibodies. Single, live, CD31-, CD45- cells were isolated by FACS for input into the chromium controller. Performed using Chromium Single Cell 3’ NextGEM Reagent Kit v3 according to the manufacturer’s instructions. Performed using Chromium Single Cell 3’ NextGEM Reagent Kit v3 according to the manufacturer’s instructions.
description
adult
developmental stage
Vcan -/-
genotype
Rot5KO
individual
Mus musculus
organism
Mus musculus
organism
heart
organism part
Mus musculus
scientific_name
male
sex
B6:Gt(ROSA)26Sortm9(cre/ESR1)Arte:Vcan-e4fl/fl
strain
Rot5KO
title