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

Overview

Journal Cancers (Basel)

Publisher MDPI

Specialty Oncology

Date 2021 Feb 5

PMID 33540698

Citations 16

Authors

Adrian Fernandez

Alfonso Navarro-Zapata

Adela Escudero

Nerea Matamala

Beatriz Ruz-Caracuel

Isabel Mirones

Alicia Pernas

Marta Cobo

Gema Casado

Diego Lanzarot

Carlos Rodriguez-Antolin

Maria Vela

Cristina Ferreras

Carmen Mestre

Aurora Viejo

Alejandra Leivas

Joaquin Martinez

Lucia Fernandez

Antonio Perez-Martinez

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

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Reovirus infection of tumor cells reduces the expression of NKG2D ligands, leading to impaired NK-cell cytotoxicity and functionality.

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