Effects of Vitamin C Supplementation to Bovine Peripheral Blood Mononuclear Cells on Antioxidant and Inflammatory Biomarker Status and Cell Viability with or without Lipopolysaccharide Stimulation

Abstract

This study aimed to determine the effects of vitamin C supplementation on antioxidant and inflammatory status and cell viability of bovine peripheral blood mononuclear cells. Peripheral blood mononuclear cells (PBMCs) were isolated from eighteen clinically healthy Japanese Black calves in this study. PBMCs were cultured with vitamin C (vitamin C group) and without vitamin C (control group) and stimulated with or without lipopolysaccharide (LPS). As a result, the total antioxidant capacities, which are the reducing power of components from Fe3 + to Fe 2 + , in the cell culture supernatant with or without LPS stimulation were significantly higher in the vitamin C group than those in the control group (P < 0.05, P < 0.01, respectively). Tumor necrosis factor-alpha in the cell culture supernatant with LPS stimulation was significantly higher in the control group than that in the vitamin C group (P < 0.05). The viability of cells cultured with LPS stimulation was significantly higher in the vitamin C group than that in the control group after 72 h of culture (P < 0.05). These results suggested that vitamin C is related to the antioxidant and anti-inflammatory properties and cell viability of PBMCs obtained from calves.

1,25‐Dihydroxyvitamin D3 potentiates the innate immune response of peripheral blood mononuclear cells from Japanese Black cattle

“…PBMCs were purified as previously described (Maeda et al, 2011;Otomaru et al, 2022Otomaru et al, , 2023. Briefly, 10 ml of heparinized blood was diluted twice in phosphate-buffered saline (PBS), placed above the separation medium solution (Lymphocyte Separation Medium 1077, Immuno-Biological Laboratories, Japan; specific gravity 1.077), and then centrifuged at 400Âg for 60 min at 15 C. Mononuclear cells at the interface were collected and then washed twice with PBS by centrifugation at 400Âg for 5 min.…”