![]() ![]() autoMACS® Pro for automated cell separation.Regenerative medicine – from basic to clinical research.Immuno-oncology – from basic research to innovative therapies.IL-2 activated NK cell immunotherapy of three children after haploidentical stem cell transplantation. Koehl U., Sorensen J., Esser R., Zimmermann S., Gruttner H.P., Tonn T., Seidl C., Seifried E., Klingebiel T., Schwabe D. Expansion of human NK-22 cells with IL-7, IL-2, and IL-1beta reveals intrinsic functional plasticity. Natural killer cell–directed therapies: Moving from unexpected results to successful strategies. Terme M., Ullrich E., Delahaye N.F., Chaput N., Zitvogel L. Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer. Miller J.S., Soignier Y., Panoskaltsis-Mortari A., McNearney S.A., Yun G.H., Fautsch S.K., McKenna D., Le C., Defor T.E., Burns L.J., et al. Deep red line: basal NK cells, green line: IL15-stimulated PBMC-derived NKAEs, yellow line: IL15-stimulated CD45RA-derived NKAEs, light blue line: IL21-stimulated CD45RA-derived NKAEs, emerald green line: IL21-stimulated PBMC-derived NKAEs. Each row in the heatmap represents a gene and each column, an individual sample. Heat map colors correspond to gene expression as indicated in the color key: red (overexpressed) and blue (underexpressed). ( D) Heat map of normalized expression values for 48 genes that are differentially expressed between PBMC and CD45RA +-derived NKAE cells. ( C) Heat map of normalized expression values for 609 genes that are differentially expressed between IL21 and IL15-stimulated NKAE cells. ( B) Heat map of normalized expression values for 2185 genes that are differentially expressed between basal NK cells and NKAE cell products. ( A) Unsupervised hierarchical clustering of NK cells using the 15,919 genes remaining after filtering. Gene expression profiles of basal NK cells and NKAE cell products. GMP-grade NK cells for clinical use can be obtained by using different starting cells and aAPC.ĬliniMACS Prodigy NK cell activation and expansion NK cell immunotherapy NKAE cells clinical-grade manufacturing. 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. 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. 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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