Fig. 4: Differentiation of gene-edited human primary B cells into LLPCs in vitro. | Cell Death & Disease

Fig. 4: Differentiation of gene-edited human primary B cells into LLPCs in vitro.

From: Engineering of α-PD-1 antibody-expressing long-lived plasma cells by CRISPR/Cas9-mediated targeted gene integration

Fig. 4

a Schematic representation of engineered B-cell differentiation into LLPCs in vitro using a multi-step cytokine culture system. b As described in a, engineered B cells differentiated into prePBs, PBs, PCs, and LLPCs respectively at indicated step. CD20, CD38, and CD138 staining were used for phenotype identification by FACS analysis at day 5, 8, 11, and 30 post gene-editing. c Proportion of plasmablasts and plasma cells at the end of the step are shown. d To confirm LLPCs phenotype, relative markers including CD27, ki67, extracellular IgG, and intracellular IgG were detected by FACS analysis. e Transcriptional signatures involved in LLPCs differentiation were tested by Quantitative real-time PCR. f The concentrations of α-PD-1 mAb in the supernatants were monitored at indicated time points during the differentiation process. Results are combined from three independent donors. The results in panels c, e and f are presented as mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001; two-tailed Student’s t test (e) and one-way ANOVA with Tukey’s post hoc test (f) were used.

Back to article page