Optimization of Large-Scale Expansion and Cryopreservation of Human Natural Killer Cells for Anti-Tumor Therapy.
- Author:
Bokyung MIN
1
;
Hana CHOI
;
Jung Hyun HER
;
Mi Young JUNG
;
Hyo Jin KIM
;
Mi young JUNG
;
Eun Kyoung LEE
;
Sung Yoo CHO
;
Yu Kyeong HWANG
;
Eui Cheol SHIN
Author Information
- Publication Type:Original Article
- Keywords: Natural killer cells; NK cell expansion; Allogeneic NK therapy; Cryopreservation; Cancer
- MeSH: Animals; Antibody-Dependent Cell Cytotoxicity; Carcinoma, Hepatocellular; Cell Survival; Cell- and Tissue-Based Therapy; Cryopreservation*; Freezing; Heterografts; Humans*; Interleukin-2; Killer Cells, Natural*; Methods; Mice; Mice, SCID; Muromonab-CD3; Phenotype; Rituximab
- From:Immune Network 2018;18(4):e31-
- CountryRepublic of Korea
- Language:English
- Abstract: Allogeneic natural killer (NK) cell therapy is a potential therapeutic approach for a variety of solid tumors. We established an expansion method for large-scale production of highly purified and functionally active NK cells, as well as a freezing medium for the expanded NK cells. In the present study, we assessed the effect of cryopreservation on the expanded NK cells in regards to viability, phenotype, and anti-tumor activity. NK cells were enormously expanded (about 15,000-fold expansion) with high viability and purity by stimulating CD³⁺ T cell-depleted peripheral blood mononuclear cells (PBMCs) with irradiated autologous PBMCs in the presence of IL-2 and OKT3 for 3 weeks. Cell viability was slightly reduced after freezing and thawing, but cytotoxicity and cytokine secretion were not significantly different. In a xenograft mouse model of hepatocellular carcinoma cells, cryopreserved NK cells had slightly lower anti-tumor efficacy than freshly expanded NK cells, but this was overcome by a 2-fold increased dose of cryopreserved NK cells. In vivo antibody-dependent cell cytotoxicity (ADCC) activity of cryopreserved NK cells was also demonstrated in a SCID mouse model injected with Raji cells with rituximab co-administration. Therefore, we demonstrated that expanded/frozen NK cells maintain viability, phenotype, and anti-tumor activity immediately after thawing, indicating that expanded/frozen NK cells can provide ‘ready-to-use’ cell therapy for cancer patients.
