Homo sapiens
Protocols: Tregs and CD4+ Teffs were isolated from buffy coats obtained from 3 volunteer blood donors. Treg were cultured at 37°C and were expanded in penicillin (100 U/ml) and streptomycin (100 mg/ml) containing SCGM medium (CellGro, CellGenix, Germany) supplemented with 10% heat inactivated human AB serum, IL-2 (2×103 U/mL, Aldesleukin, Chiron, USA) and beads coated with anti-CD3 and anti-CD28 antibodies (Invitrogen, USA) in a 1:0.6 cell:bead ratio. Beads were added to the cultures at day 0 and 7. Total RNA was isolated from Tregs and Teffs expanded for 14 days in previously described conditions using RNeasy Mini Kit (Qiagen, Hilden, Germany), followed by quality and quantity assessment using 2100 Bioanalyzer (Agilent, Santa Clara, USA). RNA with a RNA Integrity Number (RIN) of 7.0 or above was used for sequencing library preparation. SureSelect Strand Specific mRNA library kit (Agilent, Santa Clara, USA) starting with two rounds of polyA selection was used followed by the standard procedure. The cDNA libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Library Quantification Kit for Illumina (Kapa Biosystems, Woburn, USA) prior to cluster generation.
ENA first public
2017-12-07
ENA last update
2016-12-07
External Id
SAMEA23737918
INSDC center alias
Medical University of Gdansk Department of Biology and Pharmaceutical Botany
INSDC center name
Medical University of Gdansk Department of Biology and Pharmaceutical Botany
INSDC first public
2017-12-07T17:07:39Z
INSDC last update
2016-12-07T17:23:18Z
INSDC status
public
Submitter Id
E-MTAB-5322:Treg 22A
broker name
ArrayExpress
cell type
regulatory T cell
common name
human
growth condition
standard 37C
individual
Individual 22
organism part
blood
sample name
E-MTAB-5322:Treg 22A