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. 2021 Mar 3;21:107–120. doi: 10.1016/j.omtm.2021.02.023

Figure 2.

Figure 2

Antigen-dependent enrichment and expansion of NKG2D CAR-T population with K562 feeder cells

(A) Timeline for OKT3 activation, piggyBac-mediated genetic modification, and K562-based antigen-dependent expansion of NKG2D CAR-T cells. (B) Representative flow cytometry quadrant plots to illustrate the time course of antigen-dependent K562-based enrichment of NKG2D CAR-T cells from a single donor. CAR-T cells were analyzed through co-staining with anti-CD3 and anti-Streptavidin II tag antibodies. (C) Antigen-dependent enrichment of NKG2D CAR-T cells from day 7 to day 28 was similarly analyzed by flow cytometry (n = 7). (D) Antigen-dependent expansion of NKG2D CAR-T cells with K562 feeder cells from day 14 to day 28 was analyzed by trypan blue exclusion assay (n = 7). (E) ELISpot-IFNγ assay against CAOV3 and HCT-116 demonstrates that the total number of IFNγ-secreting spots increased with increasing enrichment of NKG2D CAR-T cells from day 7 to day 28. Data from 28-day Ctrl CAR-T cells are included as a negative control. Data shown are mean ± SD from a single representative assay. All data were derived from healthy donors.