RT Journal Article
T1 Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy
A1 Fernández, Adrián
A1 Navarro Zapata, Alfonso
A1 Escudero, Adela
A1 Matamala, Nerea
A1 Ruz Caracuel, Beatriz
A1 Mirones, Isabel
A1 Pernas, Alicia
A1 Cobo, Marta
A1 Casado, Gema
A1 Lanzarot, Diego
A1 Rodríguez Antolín, Carlos
A1 Vela, María
A1 Ferreras, Cristina
A1 Mestre, Carmen
A1 Viejo, Aurora
A1 Leivas, Alejandra
A1 Martínez, Joaquín
A1 Fernández, Lucía
A1 Pérez Martínez, Antonio
AB 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.
PB MDPI
SN 2072-6694
YR 2021
FD 2021-02-02
LK https://hdl.handle.net/20.500.14352/7352
UL https://hdl.handle.net/20.500.14352/7352
LA eng
NO his 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.
NO Unión Europea
NO Instituto de Salud Carlos III
NO Fundación CRIS para Vencer al Cáncer
DS Docta Complutense
RD 13 abr 2025