Sample 3
Canis lupus familiaris
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.
ENA first public
2025-10-02
INSDC center name
Department of Companion Animals Clinical Sciences, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
INSDC status
public
Submitter Id
E-MTAB-15208:Sample 3
age
8
broker name
ArrayExpress
cell type
lung cell
collection date
not collected
common name
dog
developmental stage
adult
disease
respiratory system cancer
genotype
wild type genotype
geographic location (country and/or sea)
not collected
individual
PAC3
isolate
not applicable
organism part
lung
sample name
E-MTAB-15208:Sample 3
scientific_name
Canis lupus familiaris
sex
female
strain
Cocker