characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-01T00:20:36Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-01T00:20:36Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112898432
characteristics.characteristic: External Id
characteristics.text: Gustave Roussy Cancer Campus, Villejuif Cedex, France
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-01T00:20:36Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-01T00:20:36Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14904569
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12846:mLN_Eclostri_VDJ
characteristics.characteristic: Submitter Id
characteristics.text: 6 to 7
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: T cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: CFSE+ cells were collected from tumor, tumor draining lymph node (tdLN), contralateral lymph node (cLN) and mesenteric lymph nodes (mLN), 24H after CFSE injection in mLN, in MCA205 tumor-bearing mice, colonized (E.clostri) or not (SPF) by Enterocloster clostridioformis bacteria. CFSE+ CD4 and CD8 T cells were collected with cell sorter (cytometry). Tumors, mesenteric lymph nodes (mLN), draining lymph nodes (tdLN) and contralateral lymph nodes (cLN) were harvested 24h after CFSE injection in the mLN, 10 days after tumor inoculation. Excised tumors were cut into small pieces and digested in RPMI medium containing LiberaseTM at 25 μg/ml (Roche) and DNase1 at 150 IU/ml (Roche) for 30 minutes at 37°C and then crushed and filtered twice using 100- and 70-μm cell strainers (Becton & Dickinson). Lymph nodes were crushed in RPMI medium and subsequently filtered twice through a 100-μm cell strainer. MCA205 were inoculated subcutaneously at a concentration of 0.8x10^6 cells/mL in 100µl. Tumor growth lasted 10 days. Mice were oral with water or E. clostridioformis at day 5 and day 7 after tumor inoculation. Single cell suspensions from a total of 500 to 20,000 cells with barcoded gel beads and partitioning oil were loaded to Chromium Next GEM Chip K to generate single-cell gel bead-in-emulsion. Full-length cDNA along with cell barcode identifiers were PCR-amplified to generate 5’ Gene Expression (GEX) libraries and V(D)J libraries. Libraries construction were performed using Chromium Next GEM Single Cell 5' kits v2 (PN-1000263), Chromium Next GEM Chip K, Single Cell VDJ 5' Gel Bead, Chromium Single Cell Mouse TCR amplification kit (16 rxns (reactions)) PN-1000254, Library construction kit (16 rxns) PN-1000190 and Dual Index kit TT set A (96 rxns) PN-1000215 kits according to the manufacturer’s protocol.
characteristics.characteristic: description
characteristics.text: fibrosarcoma
characteristics.characteristic: disease
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: mesenteric lymph node
characteristics.characteristic: organism part
characteristics.characteristic: sampling site
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: mLN_Eclostri_VDJ
characteristics.characteristic: title
characteristics.text: MCA205 fibrosarcoma (0.8x10^6 cells) incoculated subcutaneously
characteristics.characteristic: xenograft