CIDP_01DenStim2YY_G
Homo sapiens
Protocols: Plasmacytoid dendritic cells (pDCs) were purchased as pre-isolated cells (STEMCELL Technologies, #70046). Peripheral blood was collected using acid-citrate-dextrose solution A (ACDA) as the anticoagulant (provided by STEMCELL Technologies).. Isolated pDCs were cultured in ImmunoCult™-XF Medium (STEMCELL Technologies, #10981). Cells were maintained at 37 °C in a humidified incubator with 5% CO₂.  Stimulation with CpG oligonucleotides (InvivoGen, #tlrl-kit9h) was performed on day 1 post isolation at a final working concentration of 6 μg/mL for 24 hours prior to downstream analyses. Primary human plasmacytoid dendritic cells (pDCs) were isolated from peripheral blood (PB) mononuclear cells (MNCs) using negative immunomagnetic separation techniques (provided by STEMCELL Technologies).  Primary human plasmacytoid dendritic cells (pDCs) were isolated from peripheral blood (PB) mononuclear cells (MNCs) using negative immunomagnetic separation techniques. PB was collected using acid-citrate-dextrose solution A (ACDA) as the anticoagulant. Single-cell suspensions were prepared in advance and kept at 4 °C or on ice throughout barcoding. To enable multiplexing, cells were labeled with unique DNA barcodes using Anchor and Co-Anchor oligonucleotides (5 μM each in 1% BSA/PBS). Up to 0.5 million cells per sample were processed. Cells were washed in 2% BSA/PBS, pelleted, and resuspended in 1% BSA/PBS. Each sample was incubated sequentially on ice with 10× Anchor:Barcode solution (20 μL) followed by 10× Co-Anchor solution (20 μL), with gentle pipette mixing and 5-minute incubations between steps. After labeling, cells were washed once in 1% BSA/PBS, pelleted, and pooled in equal cell numbers. Pooling was performed by serially combining samples with 350 μL of PBS/BSA per transfer to maximize recovery. The pooled suspension was centrifuged, and cells were resuspended in a minimal volume of buffer for counting and loading into the 10x Genomics Chromium platform for 3′ gene expression profiling. The pooled, barcoded single-cell suspension was then processed using the 10x Genomics Chromium platform following the manufacturer's protocol for 3′ gene expression library preparation via Single Cell 3' v4 (polyA).
ENA first public
2026-05-23
INSDC center name
CSCI
INSDC status
public
Submitter Id
E-MTAB-16728:CIDP_01DenStim2YY_G
age
pooled
broker name
ArrayExpress
cell type
blood cell
collection date
not collected
common name
human
developmental stage
adult
disease
normal
geographic location (country and/or sea)
not collected
individual
pooled
isolate
not applicable
organism part
blood
pool id
mixed, see sample_meta.csv
sample name
E-MTAB-16728:CIDP_01DenStim2YY_G
scientific_name
Homo sapiens
sex
pooled
stimulus
mixed