2024-06-01
ENA first public
ERC000011
ENA-CHECKLIST
UCL
INSDC center name
public
INSDC status
ERS20170135
SRA accession
E-MTAB-11528_2:FL15_abTCR
Submitter Id
12+12+12+20+20+20pcw
week
age
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: Lung tissue for this project was obtained from the MRC-Wellcome Trust Human Developmental Biology Resource (https://www.hdbr.org). The HDBR obtain human embryonic and fetal samples from medical or surgical terminations of pregnancy. We requested lung tissue samples from fetuses aged between 8 and 20 post-conception weeks (pcw). We received whole lungs or lung fragments in refrigerated Hibernate E medium. Any trachea or non-lung tissue was removed by dissection. To enrich lung samples for CD45+ immune cells, we utilised fluorescence activated cell sorting (FACS). Cells were first blocked, to prevent non-specific antibody binding, using Human TruStain FcX (Biolegend, 422301) and then stained for 30 minutes on ice, using flurochrome-conjugated antibodies, CD45-PE, CD31-BV421, EpCAM-APC and CD235a-FITC. After staining, cells were resuspended in buffer containing DAPI, to enable the determination of live versus dead cells. FACS was performed on a BD FACS Aria. Appropriate ‘fluorescence minus one’ controls were used to determine FACS gating. Samples were gated to allow the collection of live single cells that were negative for CD235a (red blood cells), CD31 (endothelial cells) and EpCAM (epithelial cells), and positive for CD45 (immune cells). CD45+ immune cells were frozen prior to CITE-Seq or scRNAseq. Nucleic acid was extracted according to the standard protocol of the Chromium single cell 5'V1 kit. abTCR or BCR enrichment
description
embryo stage
developmental stage
normal
disease
not collected
geographic location (country and/or sea)
growth condition
HDBR 14732+ 14776+14794+14787+14806+15024
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
lung
organism part
abTCR
protocol
Homo sapiens
scientific_name
Mixed
sex
FL15_abTCR
title