2024-10-30
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA11327081
External Id
Instituto de Medicina Molecular - Joao Lobo Antunes, FMUL, University of Lisbon
INSDC center name
2021-11-29T16:51:28Z
INSDC last update
public
INSDC status
ERS8972309
SRA accession
E-MTAB-11211:Sample 6
Submitter Id
26.5
month
age
ArrayExpress
broker name
CD4-positive, alpha-beta thymocyte
cell_type
human
common name
Protocols: Thymocytes were isolated by Ficoll-Hypaque (GE Healthcare) from the cell suspensions obtained by thymic tissue manual dispersion. CD4SP Tregs and Tconvs were sort-purified based on the surface expression of CD4, CD8, CD27, CD25 and CD127 in FACS Aria III (BD Biosciences), reaching purities always above 95%. CD3 was intentionally not used to avoid possible signalling, but the sorting strategy was always further validated in parallel using CD3 and intracellular Foxp3 in a Fortessa flow cytometer (BD Biosciences). The staining protocols have been previously described (Nunes-Cabaço et al., 2010). Thymic samples were obtained during paediatric reconstructive cardiac surgery, using tissue that would be otherwise discarded from children without evidence of immunodeficiency or syndromatic diseases. Conversion between annotations was made with “biomaRt”. RNA samples were extracted from cell pellets of sorted 600,000 tTregs and tTconvs, using the AllPrep DNA/RNA kit (QIAGEN) and following the manufacturer’s instructions. Libraries were built selecting for polyadenylated RNA after depleting ribosomal fraction.
description
infant
developmental stage
normal
disease
T277
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
thymus
organism part
Homo sapiens
scientific_name
male
sex
Sample 6
title