Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy

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 an...

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Detalles Bibliográficos
Autores: 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, Antonio
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/7352
Acceso en línea:https://hdl.handle.net/20.500.14352/7352
Access Level:acceso abierto
Palabra clave:616-006
616.15
NK cell immunotherapy
NK cell activation and expansion
NKAE cells
Clinical-grade manufacturing
CliniMACS Prodigy
Medicina
Hematología
Oncología
32 Ciencias Médicas
3205.04 Hematología
3201.01 Oncología
Descripción
Sumario: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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