Evaluation of the effects of dimethylsulphoxide on morphology, cellular viability, mRNA, and protein expression of stem cells culture in growth media

评估二甲基亚砜对生长培养基中干细胞培养的形态、细胞活力、mRNA 和蛋白质表达的影响

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作者:Hyunjin Lee, Jun-Beom Park

Abstract

The aim of this study was to evaluate the effects of differential concentration of dimethylsulphoxide (DMSO) on the morphology, cell viability, mRNA, and protein expression of stem cells obtained from the intraoral area. Stem cells derived obtained from gingiva were cultured in a growth medium in the presence of DMSO at concentrations ranging from 0.01 to 10%. The morphology and cellular viability were evaluated on days 1, 3, 5, 7 and 10. Quantitative polymerase chain reaction was used to evaluate the mRNA levels of collagen I and Runt-related transcription factor 2 (Runx2). Immunofluorescent assays were performed for Runx2 and collagen I, and protein expressions were measured, including those of Runx2 and collagen I using western blot analysis. Cells in the control group showed normal fibroblast morphology in the growth media. Cells from the higher DMSO concentration were significantly different compared to the control. The decrease in cell viability was noted in the higher concentration. A notable change in collagen I expression was noted at the higher concentrations of DMSO groups. Based on these findings, it was concluded that DMSO may have detrimental effects on the cell morphology and viability of mesenchymal stem cells. The results also suggest that DMSO has toxic effects via reduced collagen I expression.

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