Influence of murine mesenchymal stem cells on proliferation, phenotype, vitality, and cytotoxicity of murine cytokine-induced killer cells in coculture

• Published in: PloS one Vol. 9; no. 2; p. e88115
• Main Authors: Bach, Martin, Schimmelpfennig, Christoph, Stolzing, Alexandra
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 06.02.2014; Public Library of Science (PLoS)
• Subjects: Animals; Antigen presentation; Antigens; B cells; Biocompatibility; Biology; Bone marrow; CD25 antigen; CD3 antigen; CD69 antigen; Cell Adhesion - drug effects; Cell culture; Cell Death - drug effects; Cell Line, Tumor; Cell proliferation; Cell Proliferation - drug effects; Cell surface; Cell Survival - drug effects; Cells, Cultured; Coculture Techniques; Culture Media, Conditioned - pharmacology; Cytokine-Induced Killer Cells - cytology; Cytokine-Induced Killer Cells - drug effects; Cytokines; Cytotoxicity; Dendritic cells; Genetic aspects; Genotype & phenotype; Hematopoietic stem cell transplantation; Hematopoietic stem cells; Identification; Immunology; Immunomodulation; Interferon; Interleukin; Interleukin 2; Interleukins; Killer cells; Lymphocytes; Lymphocytes T; Medical treatment; Medicine; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - drug effects; Mesenchyme; Mice, Inbred C57BL; Natural killer cells; Phenotype; Skin & tissue grafts; Staining and Labeling; Stem cell transplantation; Stem cells; Surface antigens; Therapeutic applications; Tissue engineering; Toxicity; Transplants & implants; Tumors; γ-Interferon
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0088115

Stimulating lymphocytes with Ifn-γ, anti-CD3, and interleukin-2 promotes the proliferation of a cell population coexpressing T-lymphocyte surface antigens such as CD3, CD8a, and CD25 as well as natural killer cell markers such as NK1.1, CD49, and CD69. These cells, referred to as cytokine-induced killer cells (CIKs), display cytotoxic activity against tumour cells, even without prior antigen presentation, and offer a new cell-based approach to the treatment of malignant diseases. Because CIKs are limited in vivo, strategies to optimize in vitro culture yield are required. In the last 10 years, mesenchymal stem cells (MSCs) have gathered considerable attention. Aside from their uses in tissue engineering and as support in haematopoietic stem cell transplantations, MSCs show notable immunomodulatory characteristics, providing further possibilities for therapeutic applications. In this study, we investigated the influence of murine MSCs on proliferation, phenotype, vitality, and cytotoxicity of murine CIKs in a coculture system. We found that CIKs in coculture proliferated within 7 days, with an average growth factor of 18.84, whereas controls grew with an average factor of 3.7 in the same period. Furthermore, higher vitality was noted in cocultured CIKs than in controls. Cell phenotype was unaffected by coculture with MSCs and, notably, coculture did not impact cytotoxicity against the tumour cells analysed. The findings suggest that cell-cell contact is primarily responsible for these effects. Humoral interactions play only a minor role. Furthermore, no phenotypical MSCs were detected after coculture for 4 h, suggesting the occurrence of immune reactions between CIKs and MSCs. Further investigations with DiD-labelled MSCs revealed that the observed disappearance of MSCs appears not to be due to differentiation processes.