Protocols: Ccl19-Cre x R26R-EYFP mice were subcutaneously injected in one flank with 5 mg of LPS from E.coli (Sigma) together with 100 mg Ovalbumin grade VI (Sigma). Mice were sacrificed at day 3 post administration and brachial lymph nodes were removed for further lymph node stromal cell isolation and scRNA sequencing analysis. Brachial lymph nodes were transferred into a 24-well dish filled with RPMI 1640 medium containing 2% FCS, 20 mM Hepes (all from Lonza), 1 mg/ml Collagenase D (Sigma), 25 µg/ml DNAse I (Applichem) and Dispase (Roche). Dissociated tissues were incubated at 37°C for 30 minutes. After enzymatic digestion, cell suspensions were washed with PBS containing 0.5% FCS and 10 mM EDTA. Stromal cells were enriched by depleting CD45+ hematopoietic cells and TER119+ erythrocytes using MACS microbeads (Miltenyi, Germany). Cells were sorted for EYFP+CD31-CD45- reticular cells and processed using the 10x Chromium (10X Genomics) system. cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3' Reagent Kit (v3 Chemistry).
ENA-FIRST-PUBLIC
2022-08-31
ENA-LAST-UPDATE
2022-08-31
External Id
SAMEA9908356
INSDC center alias
Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
INSDC center name
Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
INSDC first public
2022-08-31T00:12:58Z
INSDC last update
2022-08-31T00:12:58Z
INSDC status
public
Submitter Id
E-MTAB-10908:Sample 3
age
8
broker name
ArrayExpress
cell_type
fibroblast
common name
house mouse
developmental stage
adult
genotype
C57BL/6N-Tg(Ccl19-Cre) x R26-EYFP
immunophenotype
EYFP+CD31-CD45-
individual
1
organism part
brachial lymph node
sample name
E-MTAB-10908:Sample 3
sex
female
strain
C57Bl/6N