2025-10-02
ENA first public
ERC000011
ENA-CHECKLIST
Department of Companion Animals Clinical Sciences, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
INSDC center name
public
INSDC status
ERS24832631
SRA accession
E-MTAB-15208:Sample 3
Submitter Id
8
year
age
ArrayExpress
broker name
lung cell
cell type
not collected
collection date
dog
common name
Protocols: Biopsies from three solitary canine primary PAC were collected immediately after complete lobectomy in client-owned dogs, with informed owner consent. In each tumour, two adjacent biopsies were obtained at the tumour edge: one was fixed in 10% neutral buffered formalin and embedded in paraffin for histopathological confirmation of diagnosis, and one was transported in HBSS (Gibco) containing 5% v/v of FBS (Gibco) on ice for immediate processing. Each lung sample underwent mechanical dissociation with razor blades and was suspended in HBSS + 5% FBS with collagenase A (1 mg/mL; Sigma) and DNase I (0.05 mg/mL; Roche) before incubation at 37°C for 45 min. The cells were then filtered through a 70 µm cell strainer (BD Falcon) and resuspended in PBS (Biowest) containing 10 mM EDTA (Merck Millipore). Red blood cells were lysed as needed with a lysis buffer containing 0.15 M NH4Cl, 0.01 M KHCO3 and 0.1 mM EDTA at pH 7.5. The final cell suspension contained between 500 and 1000 cells/µL in PBS containing 0.04% of BSA (Sigma) and 0.2 U/µL of RNase inhibitor (Roche). Final cell viability was assessed by trypan blue staining and considered acceptable above 70%. Approximatively 10,000 cells from each lung sample were loaded in a Chromium Controller or Chromium iX instrument (10x Genomics, Pleasanton, CA) and encapsuled with unique barcoded primers using the drop-sequencing method according to manufacturer’s instructions Emulsion breakage, cDNA amplification and libraries construction were performed using Chromium Single Cell 3′ reagent kit v2 (10x Genomics) according to manufacturer’s instructions.
description
adult
developmental stage
respiratory system cancer
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
PAC3
individual
not applicable
isolate
Canis lupus familiaris
organism
Canis lupus familiaris
organism
lung
organism part
Canis lupus familiaris
scientific_name
female
sex
Cocker
strain
Sample 3
title