2023-05-01
ENA-FIRST-PUBLIC
2023-05-01
ENA-LAST-UPDATE
SAMN34586592
External Id
SAMEA112192601
External Id
European Bioinformatics Institute
INSDC center name
2023-05-20T18:40:32.117Z
INSDC first public
2023-05-20T18:40:32.117Z
INSDC last update
SRS17537858
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
SRS17537858
SRA accession
6 to 10
age
fibroblast
cell_type
house mouse
common name
Protocols: Mice were sacrificed at age 6-10 weeks and lymph nodes, spleen and Peyer's patches were removed for further stromal cell or immune cell isolation and scRNA sequencing analysis. For cell preparation tissues were cut into small pieces. For enzymatic digestion, cut tissues from murine lymph nodes, spleen and Peyer's patches were collected in RPMI 1640 medium containing 2% FCS, 20 mM HEPES pH 7.2 (Lonza), 0.375 mg/ml Collagenase P (Roche) and 25 mg/ml DNaseI (Applichem). Dissociated tissue was incubated at 37°C for 60 min, with resuspension and collection of the supernatant every 15 min. For cell-sorting experiments, stromal cells were enriched by incubating the cell suspension with MACS anti-CD45 and anti-TER119 microbeads (Miltenyi Biotec) and passing it through a MACS LS column (Miltenyi Biotec). The unbound single cell suspension was stained for cell sorting with a BD FACS Melody cell sorter (BD Biosciences) and run on the 10x Chromium analyzer (10X Genomics). For isolation of hematopoietic cells from murine organs, tissues were gently smashed across a 26-gauge wire mesh and washed with medium (RMPI-1640 containing 2% FCS (Sigma), 1% Penicillin-Streptomycin (Corning) and 20mM HEPES (Sigma)) until further staining for cell sorting. cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3' Reagent Kit (v3 Chemistry). VSV immunization protocol: Mice were immunized intravenously with 2 × 107 plaque-forming units (PFU) of VSV. Draining inguinal lymph nodes and spleens were collected on day 14 after immunization for further processing for single cell RNA-sequencing.
description
adult
developmental stage
Tg(Cxcl13-Cre/TdTomato)+ x R26-EYFP++
genotype
mixed pool of 8 to 12 individuals
individual
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
Mus musculus
organism
Peyer's patch
organism part
Mus musculus
scientific_name
mixed
sex
C57Bl/6N
strain
Sample 19
title