Fig. 5

Hyperosmosis facilitates the efficient production of functional HSC-CAR-NK cells. A Workflow detailing the production of CAR19 NK cells via HSC differentiation. B Flow cytometry assessment of the proportion of BFP-positive cells of PB-NK cells following a 4-day culture and HSPCs following an 8-day culture or NK cell differentiation process, post-transduction with a CAR19-BFP lentiviral vector. The percentages indicate the fraction of BFP-positive cells. C Assessment of the fraction of CAR19-BFP positive cells following lentiviral transduction in PB-NK cells, HSPCs as demonstrated in (B), and differentiated HSPCs (D0 HSC-NK) bulk populations (bulk cells). D Schematic of the co-culture setup of expanded HSC-NK cells with GFP⁺ Raji cells. E Proportion of BFP⁺ NK cells and GFP⁺ Raji cells as depicted in (D). F Flow cytometry-evaluated killing activity against CD19⁺ GFP⁺ Raji cells after a 24-h co-culture of HSC-BFP-NK cells or HSC-CAR19-NK cells and Raji cells at the indicated E:T ratios. Data in C, E and F are represented as mean ± SEM from at least 3 independent experimental replicates. P values calculated by a paired Student's t-test are provided in (C), whereas P values computed by a two-way ANOVA test are indicated in (E) and (F)