2025-12-07
ENA first public
ERC000011
ENA-CHECKLIST
iTeos Therapeutics
INSDC center name
public
INSDC status
ERS25339729
SRA accession
E-MTAB-15419:GC137775
Submitter Id
ArrayExpress
broker name
B cell
cell type
not collected
collection date
human
common name
Protocols: Tonsil sample collected by adult patients undergoing tonsillectomy was cut into small fragments before mechanical dissociation through a 100 mm cell strainer using a syringe plunger. Tonsillar mononuclear cells (TMNC) were then isolated by lymphoprep density gradient centrifugation using SepMate 50 tubes (STEMCELL Technologies) and tonsillar B cells were enriched using EasySep™ Human Pan-B Cell Enrichment Kit (STEMCELL Technologies) following the manufacturer’s protocol. Isolated B cells were cultured in a 24-well plate at a concentration of 2 million per well in 1ml of X-vivo medium supplemented with 5% inactivated human serum, 1mM of Na-pyruvate and 1mg/ml of CpG for 3h at 37°C, 5% CO2. The A2AR agonist CGS-21680 was added at 5 mM alone or in combination with the A2AR antagonist inupadenant (EOS100850) at 300 nM final concentration. Treated cells were then fixed 1h at room temperature following the Fixation of Cells & Nuclei for Chromium Fixed RNA Profiling (10X Genomics) manufacturer’s protocol. Library preparation was performed following the Chromium FixedRNAProfiling_MULTIPLEXED Samples (10X Genomics) protocol. Chromium Fixed RNA Kit, Human Transcriptome, 4 rxns x 16 BC, PN-1000476
description
adult
developmental stage
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
tonsil
organism part
Homo sapiens
scientific_name
CpG
stimulus
GC137775
title