2025-11-27
ENA first public
ERC000011
ENA-CHECKLIST
Omiics, Aarhus, Denmark
INSDC center name
public
INSDC status
ERS27177076
SRA accession
E-MTAB-15969:PD1.hi.cells_2
Submitter Id
ArrayExpress
broker name
fibroblast
cell type
not collected
collection date
human
common name
Protocols: Fibroblasts from HC skin in passage 3 were thawed and cultured in a T175 flask (Sarstedt, cat. no. 83.3911.002) in DMEM low glucose media (VWR, cat. no. 392-0407). When confluency was reached cells were trypsinized and seeded into a 96-well plate and stimulated by adding 20% supernatant from PD-1+/- cultures (n=3), IFNγ (10 µg/ml) (n=3), PD-1+/- T cells (n=3), or no stimulation (n=3). After 24 hours T cells were removed. RNA was purified from the co-cultured fibroblasts after washing with PBS and cell lysis. RNA-purification was done using the RNeasy Mini Kit (Qiagen, cat. no. 74104) (n=1 in triplicates), and the Genomic DNA was removed by using the RNase-Free DNase Set (Qiagen, cat. no. 79254).  The RNA integrity number and concentration were measured on Bioanalyzer RNA Nano chip (Agilent, cat.no. 5067-1511). The SMARTer Stranded Total RNA Sample Prep Kit - HI Mammalian (Takara, Japan, cat.no. 634876) was used for library preparation with 200ng of the purified RNA as input following the manufacturer`s protocol. The Bioanalyzer High sensitivity DNA analysis kit (Agilent, cat.no. 5067-4626) was used to determine the size of the library fragments and the Qubit dsDNA High Sensitivity Kit (Invitrogen, cat. no. Q32851) was used to quantify the library.
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
Homo sapiens
scientific_name
co-culture with PD-1 hi T cell
stimulus
PD1.hi.cells_2
title