A serum-free protocol for the ex vivo expansion of Cytokine-Induced Killer cells using gas-permeable static culture flasks

Pierangela Palmerini, Anna Dalla Pietà, Roberta Sommaggio, Annavera Ventura, Giuseppe Astori, Katia Chieregato, Maria Chiara Tisi, Carlo Visco, Omar Perbellini, Marco Ruggeri, Elisa Cappuzzello, Antonio Rosato

Research output: Contribution to journalArticlepeer-review

Abstract

Cytokine-Induced (CIK) cells represent an attractive approach for cell-based immunotherapy, as they show several advantages compared with other strategies. Here we describe an original serum-free protocol for CIK cell expansion that employs G-Rex devices and compare the resulting growth, viability, phenotypic profile and cytotoxic activity with conventional culture in tissue flasks. CIK cells were obtained from buffy coats, seeded in parallel in G-Rex and tissue flasks, and stimulated with clinical-grade IFN-γ, anti-CD3 antibody and IL-2. G-Rex led to large numbers of CIK cells, with a minimal need for technical interventions, thus reducing the time and costs of culture manipulation. CIK cells generated in G-Rex showed a less differentiated phenotype, with a significantly higher expression of naive-associated markers such as CD62L, CD45RA and CCR7, which correlates with a remarkable expansion potential in culture and could lead to longer persistence and a more sustained anti-tumor response in vivo. The described procedure can be easily translated to large-scale production under Good Manufacturing Practice. Overall, this protocol has strong advantages over existing procedures, as it allows easier, time-saving and cost-effective production of CIK effector cells, fostering their clinical application.

Original languageEnglish
Pages (from-to)511-518
Number of pages8
JournalCytotherapy
Volume22
Issue number9
DOIs
Publication statusPublished - Sep 2020

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