2024-12-09
ENA first public
ERC000011
ENA-CHECKLIST
University of Glasow - School of Infection & Immunity
INSDC center name
public
INSDC status
ERS20302519
SRA accession
E-MTAB-14198:GAU-DS-s002
Submitter Id
ArrayExpress
broker name
Mixure (see metadata plate2)
cell_type
not collected
collection date
human
common name
Protocols: DC2, DC3, and iDC3, as well as SPP1pos and CLEC10Apos STMs to serve as negative controls, were sorted using Sony MA900 directly into two 384-well plates to perform ST-DC index SORT-seq  Fresh synovial tissues were diced into 1–2-mm3 fragments with a sterile disposable no. 22 scalpel and transferred to a sterile universal container containing 10 ml of sterile RPMI with penicillin/streptomycin 100 U ml–1 and l-glutamine 2 mM (RPMI medium) in a 1:33 dilution of Liberase at 0.15μg ml–1, 0.78 Wünsch units ml–1 (TM Research Grade, Thermolysin, medium, Roche Diagnostics (no. 000000005401127001, Sigma)). Tissue pieces were incubated at 37 °C, 5% CO2 in a humidified atmosphere for 30–45 min, with rotation on a Miltenyi MACSmix tube-rotator and vigorous shaking by hand twice during this incubation. After incubation, the digested mixture was filtered using an Easy Strain 100 μM cell strainer into a 50-ml falcon tube. Residual cell clumps retained on the filter were gently massaged using the rubber end of a 1-ml syringe plunger, to optimize cell retrieval. Complete medium (RPMI with 10% fetal calf serum (FCS) was poured through the filter into the falcon tube up to 40 ml then centrifuged at 1,800 r.p.m. for 10 min at 4 °C, and the supernatant was carefully removed. One milliliter of complete medium was added to gently resuspend the cell pellet, using a wide-opening 1-ml pipette tip to minimize cell damage, then the resuspended cells were transferred to a sterile Eppendorf tube. A 20-μl aliquot was used to count the cells. Cells were centrifuged at 1,500 r.p.m. for 5 min at 4 °C. The supernatant was removed, and cells were stained for FACS sorting Nuclei acid extraction was provided by Single Cell Discoveries (Utrecht, Netherlands) Library generation was provided by Single Cell Discoveries (Utrecht, Netherlands)
description
rheumatoid arthritis
disease
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
synovial membrane
organism part
Homo sapiens
scientific_name
GAU-DS-s002
title