2025-03-31
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
INSDC center name
public
INSDC status
ERS23862136
SRA accession
E-MTAB-14987:Sample 4
Submitter Id
19
year
age
ArrayExpress
broker name
hematopoietic cell
cell type
not collected
collection date
human
common name
Protocols: After surgical excision, tonsils were collected in sterile phosphate buffered saline on ice and processed within 1 hour of resection. Heavily clotted or cauterized areas of the tissue were removed and tonsils were dissected into small pieces. For the isolation of stromal cells, tissue was digested using a MACS human tumor dissociation kit (Miltenyi Biotec) according to the manufacturer’s protocol. To enrich for stromal cells, hematopoietic and erythrocytic cells were depleted by incubating the cell suspension with MACS anti-CD45 and anti-CD235a (Glycophorin A) MicroBeads (Miltenyi Biotec) and passing it through a MACS LS column (Miltenyi Biotec). The unbound single cell suspension was stained for further cell sorting. For the isolation of hematopoietic cells, tissues were gently smashed across a 26-gauge wire mesh using a syringe plunger and washed with medium (RMPI1640 containing 2% FCS (Sigma), 1% Penicillin-Streptomycin (Corning) and 20 mM HEPES (Sigma)) until further staining for cell sorting. Hematopoietic cells were stained with fluorochrome-conjugated antibodies against human CD45, CD3, CD19 and CD14 and stromal cells with fluorochrome-conjugated antibodies against human CD45 and CD235a. Live/dead cell discrimination was performed by adding 7-amino-actinomycin D (7AAD; Calbiochem) prior to acquisition. CD45– CD235a– stromal cells, CD45+ CD3+ T cells and CD45+ CD19+ B cells were sorted with a BD FACS Melody cell sorter (BD Biosciences) and run on the 10x Chromium analyzer (10X Genomics). cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3’ Reagent Kit (v3 Chemistry).
description
adult
developmental stage
Tonsillitis
disease
not collected
geographic location (country and/or sea)
Inflamed 01
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
palatine tonsil
organism part
Homo sapiens
scientific_name
male
sex
Sample 4
title