2025-08-24
ENA first public
ERC000011
ENA-CHECKLIST
ArrayExpress
INSDC center name
public
INSDC status
ERS25357912
SRA accession
E-MTAB-15436_3:Donor 1 - GEX
Submitter Id
45
year
age
ArrayExpress
broker name
CD8-positive, alpha-beta T cell
cell type
not collected
collection date
human
common name
Protocols: Freshly isolated nasal cells and PBMCs were rested in culture medium (AIM-V supplemented with 2% AB serum) for 2 hours at 37 °C. Following the resting period, half of the cells were stimulated with anti-CD3/CD28 Dynabeads at a 1:1 cell-to-bead ratio, while the remaining half were left unstimulated as controls. Cells were cultured overnight in 96-well U-bottom tissue culture plates under standard incubation conditions (37 °C, 5% CO2). After 24 hours, both nasal and blood T cells were washed with PBS and subsequently stained with surface markers (NIR, CD3, CD4, CD8, CD45RO) and sorted using FACS (BD FACSAria III). Specifically, NIR-CD3+CD4-CD8+CD45RO+ cells were sorted from each sample for the single-cell RNA sequencing experiment. The sorted cells were labeled with distinct Hash Tag Oligonucleotides (HTOs) for multiplexing.  Single-cell suspensions were loaded onto the 10x Genomics Chromium Controller for partitioning into Gel Bead-In-Emulsion (GEMs) using the Chromium Next GEM Single Cell 5' Kit v2 (10x Genomics). Reverse transcription was performed within the GEMs, followed by amplification and library construction. HTO libraries were constructed following the TotalSeq™ protocol.  Reverse transcription was performed within the GEMs, followed by amplification and library construction. HTO libraries were constructed following the TotalSeq™ protocol.
description
adult
developmental stage
normal
disease
not collected
geographic location (country and/or sea)
Donor 1
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
nasal cavity and pbmc
organism part
Homo sapiens
scientific_name
female
sex
negative or cd3/28 beads
stimulus
Donor 1 - GEX
title