Optimizing the procedure to manufacture clinical‐grade NK cells for adoptive immunotherapy
Author
Fernández, Adrián; Navarro‐zapata, Alfonso; Escudero, Adela; Matamala, Nerea; Ruz‐caracuel, Beatriz; Mirones, Isabel; Pernas, Alicia; Cobo, Marta; Casado, Gema; Lanzarot, Diego; Rodríguez‐antolín, Carlos; Vela, María; Ferreras, Cristina; Mestre, Carmen; Viejo, Aurora; Leivas, Alejandra; Martínez, Joaquín; Fernández, Lucía; Pérez Martínez, AntonioEntity
UAM. Departamento de PediatríaPublisher
MDPIDate
2021-02-02Citation
10.3390/cancers13030577
Cancers 13.3 (2021): 577
ISSN
2072-6694DOI
10.3390/cancers13030577Funded by
This work was supported by the National Health Service of Spain, Instituto de Salud Carlos III (ISCIII), FONDOS FEDER grant (FIS) PI18/01301 to Pérez‐Martínez A, CRIS Foundation to Beat Cancer to Escudero A, Fernández A; Navarro A, Mirones I, and Fundación Mari Paz Jiménez Casado and La Sonrisa de Álex to Vela M.Project
Gobierno de España. PI18/01301Subjects
Clinical‐grade manufacturing; CliniMACS Prodigy; NK cell activation and expansion; NK cell immunotherapy; NKAE cells; MedicinaRights
© 2021 by the authorsAbstract
Natural killer (NK) cells represent promising tools for cancer immunotherapy. We report the optimization of an NK cell activation–expansion process and its validation on clinical‐scale. Methods: RPMI‐1640, stem cell growth medium (SCGM), NK MACS and TexMACS were used as culture mediums. 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. Fold increase, NK cell purity, activation status, cytotoxicity and transcriptome profile were analyzed. Clinical‐grade NKAE cells were manufactured in CliniMACS Prodigy. Results: NK MACS and TexMACs achieved the highest NK cell purity and lowest T cell contamination. Obtaining NKAE cells from CD45RA+ cells was feasible although PBMC yielded higher total cell numbers and NK cell purity than CD45RA+ cells. The highest fold expansion and NK purity were achieved by using PBMC and K562mbIL21‐41BBL cells. However, no differences in activation and cytotoxicity were found when using either NK cell source or activating cell line. Transcriptome profile showed to be different between basal NK cells and NKAE cells expanded with K562mbIL21‐41BBL or K562mbIL15‐41BBL. Clinical‐grade manufactured NKAE cells complied with the specifications from the Spanish Regulatory Agency. Conclusions: GMP‐grade NK cells for clinical use can be obtained by using different starting cells and aAPC
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Google Scholar:Fernández, Adrián
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Navarro‐zapata, Alfonso
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Escudero, Adela
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Matamala, Nerea
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Ruz‐caracuel, Beatriz
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Mirones, Isabel
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Pernas, Alicia
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Cobo, Marta
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Casado, Gema
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Lanzarot, Diego
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Rodríguez‐antolín, Carlos
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Vela, María
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Ferreras, Cristina
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Mestre, Carmen
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Viejo, Aurora
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Leivas, Alejandra
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Martínez, Joaquín
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Fernández, Lucía
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Pérez Martínez, Antonio
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