Influence of Murine Mesenchymal Stem Cells on Proliferation, Phenotype, Vitality, and Cytotoxicity of Murine Cytokine-Induced Killer Cells in Coculture

Abstract

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.

Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease

“…Transgenic mice expressing eGFP (8–12 weeks old, male, C57Bl/6-Tg(UBC-GFP)30Scha/J (eGFP mice) were obtained from Jackson Laboratories (Bar Harbor, ME). Bone marrow was harvested from tibia and femur as described previously [78]. MSCs were cultivated in minimum essential medium (MEM) α, GlutaMAX™ (Gibco, 32561029) with 15% fetal calf serum (FCS) (Gibco, 10270106) and 1% penicillin/streptomycin (Gibco, 15070-063) supplemented with 20 ng/mL FGFb (Peprotech, 450-33).…”

Distribution pattern following systemic mesenchymal stem cell injection depends on the age of the recipient and neuronal health

“…After isolation, bone marrow from tibiae and femurae were harvested as described previously [ 40 ] and cultured in DMEM 1×, low glucose (Gibco) with GlutamaxI (Gibco), 10% fetal calf serum (Biochrome), and 1% penicillin/streptomycin (Gibco) for the isolation of adherent MSCs. Briefly, mouse bone marrow cells were obtained by centrifugation from tibiae and femurae and were cultured according to the method of Dobson et al [ 41 ].…”

Influence of age on rat bone-marrow mesenchymal stem cells potential

“…Mesenchymal Stem Cells (MSCs) have self-renewal capacity and multiple differentiation potentials, and a priori , could play important roles in regenerative medicine. The promising role of MSCs in cell-based therapies is their trophic, paracrine and immunomodulatory functions that may have the greatest therapeutic impact in vivo 1 2 . Tissue engineering from MSCs are of highly importance for regeneration of mesenchymal tissues such as craniofacial bone 3 , cartilage 4 and connective tissues 5 .…”