Immune Netw.  2018 Aug;18(4):e31. 10.4110/in.2018.18.e31.

Optimization of Large-Scale Expansion and Cryopreservation of Human Natural Killer Cells for Anti-Tumor Therapy

Affiliations
  • 1BioMedical Science and Engineering Interdisciplinary Program, KAIST, Daejeon 34141, Korea. ecshin@kaist.ac.kr
  • 2Cell Therapy Research Center, GC LabCell, Yongin 16924, Korea. ykhwang@mogam.re.kr
  • 3GC Pharma, GC LabCell, Yongin 16924, Korea.
  • 4Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon 34141, Korea.

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 CD³âº 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.

Keyword

Natural killer cells; NK cell expansion; Allogeneic NK therapy; Cryopreservation; Cancer

MeSH Terms

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
Interleukin-2
Muromonab-CD3
Rituximab
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