2024-05-06
ENA first public
ERC000011
ENA-CHECKLIST
PhD student  AG Blank - Molecular and Translational Allergology Center of Allergy and Environment (ZAUM) Technical University of Munich (TUM) - School of Medicine Helmholtz Center Munich, German Research Center for Environmental Health (GmbH) - Institute of Allergy Research (IAF)
INSDC center name
public
INSDC status
ERS19061015
SRA accession
E-MTAB-14030:Sample 4
Submitter Id
5 to 8
week
age
ArrayExpress
broker name
CD4-positive helper T cell
cell_type
not collected
collection date
house mouse
common name
Protocols:  To isolate pulmonary leukocytes, lungs were removed and chopped into small pieces. Digestion of lung tissue was performed in RPMI medium (Thermo Fisher Scientific) containing 1 mg/mL collagenase A and 100 µg/mL DNAse I (both from Sigma-Aldrich, Merck, Darmstadt, Germany) for 30 min at 37° C. Digested lung tissue was mashed trough a 70 µm cell strainer. Cells were pelleted (400 g, 4 °C, 5 min), resuspended in 5 mL 40% percoll in RPMI (v/v) solution, underlayered with 5 mL 80% percoll solution, and centrifuged (1600 g, RT, 15 min). Lymphocytes were collected from the interphase and washed with PBS. CD4+ T cells were enriched from pulmonary lymphocytes with the EasySepTM Mouse CD4+ T cell Isolation Kit and the EasyEightsTM EasySepTM Magnet (both from STEMCELL Technologies, Vancouver, Canada) according to the manufacturer’s instructions.  All libraries for scRNA-seq were prepared according to the manual (CG000315 Rev A) of the Chromium Next GEM Single Cell 3’ Reagent Kit v3.1 (Dual Index) provided by 10x Genomics B.V. (Leiden, The Netherlands). A cell recovery of 10.000 was targeted. All libraries for scRNA-seq were prepared according to the manual (CG000315 Rev A) of the Chromium Next GEM Single Cell 3’ Reagent Kit v3.1 (Dual Index) provided by 10x Genomics B.V. (Leiden, The Netherlands). A cell recovery of 10.000 was targeted.
description
adult
developmental stage
allergy
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
SH17 _ 10-12 _ 31-33
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
lung
organism part
Mus musculus
scientific_name
female
sex
C57BL/6
strain
Sample 4
title