Homo sapiens
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)
ENA first public
2024-12-09
INSDC center name
University of Glasow - School of Infection & Immunity
INSDC status
public
Submitter Id
E-MTAB-14198:GAU-DS-s001
broker name
ArrayExpress
cell_type
Mixure (see metadata plate1)
collection date
not collected
common name
human
disease
rheumatoid arthritis
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
synovial membrane
sample name
E-MTAB-14198:GAU-DS-s001
scientific_name
Homo sapiens