Clinical-grade manufactured NKAE cells complied with the specifications from the Spanish Regulatory Agency. Transcriptome profile showed to be different between basal NK cells and NKAE cells expanded with K562mbIL21-41BBL or K562mbIL15-41BBL. However, no differences in activation and cytotoxicity were found when using either NK cell source or activating cell line. The highest fold expansion and NK purity were achieved by using PBMC and K562mbIL21-41BBL cells.
Obtaining NKAE cells from CD45RA + cells was feasible although PBMC yielded higher total cell numbers and NK cell purity than CD45RA + cells. Results: NK MACS and TexMACs achieved the highest NK cell purity and lowest T cell contamination. Clinical-grade NKAE cells were manufactured in CliniMACS Prodigy. Fold increase, NK cell purity, activation status, cytotoxicity and transcriptome profile were analyzed. Activated and expanded NK cells (NKAE) were obtained by coculturing total peripheral blood mononuclear cells (PBMC) or CD45RA + cells with irradiated K562mbIL15-41BBL or K562mbIL21-41BBL. Methods: RPMI-1640, stem cell growth medium (SCGM), NK MACS and TexMACS were used as culture mediums. We report the optimization of an NK cell activation–expansion process and its validation on clinical-scale. Natural killer (NK) cells represent promising tools for cancer immunotherapy.
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