2024-04-01
ENA first public
ERC000011
ENA-CHECKLIST
CARDIFF
INSDC center name
public
INSDC status
ERS18396940
SRA accession
E-MTAB-13875:Sample 15
Submitter Id
ArrayExpress
broker name
PBMC
cell_line
monocyte
cell_type
not collected
collection date
human
common name
Protocols: After three days in culture, CD3+ Vδ2+ T-cells were enriched by cell sorting using a FACSAria II (BD Biosciences), to purities >99.8%. Monocytes were freshly isolated and irradiated at 50 Gy prior to co-culture with freshly isolated autologous at a monocyte to γδ T-cell ratio of 1:10.  Cultures were treated with 10 nM HMB-PP alone, or in combination with 100 U/ml IL-2, 20 ng/ml IL-15 or 20 ng/ml IL-21.  RNA was extracted from cell pellets using the RNeasy Micro Kit (Qiagen) and examined qualitatively and quantitatively using the TapeStation 2200 system (Agilent) and a Qubit fluorometer (Thermo Fisher Scientific).  Ribosomal RNA was depleted as part of the Illumina Stranded Total RNA Prep protocol using coated magnetic beads; the remaining RNA was fragmented and denatured, and first and second strand cDNA synthesis and anchor ligation was carried out according to the manufacturer’s instructions.
description
adult
developmental stage
not collected
geographic location (country and/or sea)
irradiation and co-culture with gamma-delta T cells
growth condition
D1
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
TR02
replicate
Homo sapiens
scientific_name
AH33YJBGXY
sequencing run
Sample 15
title