organism: Mus musculus
description: 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.
attributes.tag: strain
attributes.value: C57BL/6
attributes.tag: disease
attributes.value: allergy
attributes.tag: individual
attributes.value: SH17 _ 7-9 _ 28-30
attributes.tag: organism
attributes.value: Mus musculus
attributes.tag: organism
attributes.value: Mus musculus
attributes.tag: collection date
attributes.value: not collected
attributes.tag: sex
attributes.value: female
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: common name
attributes.value: house mouse
attributes.tag: organism part
attributes.value: lung
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: cell_type
attributes.value: CD4-positive helper T cell
attributes.tag: age
attributes.value: 5 to 8
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected