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Single cell RNAseq analysis of cultured human B cell

We developed an optimized in vitro model that drives human naïve B cells into ASCs through a two-step culture on stromal feeders. In the first phase, B cells were activated on CD40L-expressing stromal cells with BAFF, IL-4, IL-21, and IL-2; in the second, they were transferred to MS5 fibroblasts with IL-6 to promote terminal differentiation confirmed by the generation of CD20loCD38hiCD138+ plasma cells. To map transcriptional dynamics, we performed scRNA-seq at day 0 (D0), D6, D8, and D10, analyzing 17,220 cells that segregated into 12 clusters

Type: RNASeq
Archive: European Genome-phenome Archive (EGA)

1 Dataset

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Dataset ID	Description	Technology	Samples
EGAD50000002118	Tonsils B Cell from three different donors were cultured on our in vitro differentiation model. A mean of 20,000 viable human B cells per donor were hashed with cell-surface oligo (HTO) barcodes and pooled at days 0, 6, 8, and 10 of culture. Single-cell 5’ gene expression libraries and V(D)J enriched libraries were performed using Genomics Chromium Single Cell Immune Profiling Solution (10x Genomics) according to the standard manufacturer’s protocol. Libraries were sequenced on an Illumina NovaSeq 6000 (2 × 150 bp) with an average depth of ≥50,000 reads per cell.	Illumina NovaSeq 6000	4