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

Lee, Hyunjin; Park, Jun‐Beom

doi:10.3892/br.2017.961

Cited by 19 publications

(14 citation statements)

References 17 publications

(21 reference statements)

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“…Preliminary study to set up the final DMSO concentration for a selected cell type is needed at the start of each experiment. In both plant and animal science, the range of 0.5-1% DMSO is being applied to cells, and its final concentration 3% or higher is not advisable (Lee and Park, 2017). In different animal cell lines, it has also been suggested to limit the concentration to a maximum of 1% not only considering its toxicity for the cell lines but also because it can alter a plethora of cell responses (Hollebeeck et al, 2011).…”

Section: Resultsmentioning

confidence: 99%

Reactive Oxygen Species Imaging in U937 Cells

et al. 2020

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The U937 cell culture is a pro-monocytic, human histiocytic lymphoma cell line. These monocytes can differentiate into either macrophages or dendritic cells (antigenpresenting cells) depending on the initiators. The U937 cells activated in the presence of phorbol 12-myristate 13-acetate (PMA) change their morphology into macrophagelike cells creating pseudopodia and adhering generously. Macrophages are known to produce reactive oxygen species (ROS) mostly during phagocytosis of foreign particles, an important non-specific immune response. Recently, we have focused on the role of hydroxyl radical (HO •) and provide evidence on its importance for differentiation in U937 cells. Based on electron paramagnetic resonance (EPR) spectroscopy combined with confocal laser scanning microscopy (CLSM), formation of HO • was confirmed within the cells undergoing differentiation and/or apoptosis during the PMA treatment. This study aims to increase our knowledge of ROS metabolism in model cell lines used in human research.

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Section: Resultsmentioning

confidence: 99%

Reactive Oxygen Species Imaging in U937 Cells

et al. 2020

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“…Furthermore, DMSO may have detrimental effects on cell morphology (13). In our study, morphological changes of WM-266-4 cells exposed to 1-5% DMSO were observed.…”

Section: Resultsmentioning

confidence: 46%

The influence of dimethyl sulfoxide (DMSO) on metabolic activity and morphology of melanoma cell line WM-266-4

Ferk

Dariš

2018

Cell Mol Biol (Noisy-le-grand)

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Dimethyl sulphoxide (DMSO) is widely used as a solvent in biomedical research, regularly in concentrations up to 1%. Nevertheless, little is known about the effect of different DMSO concentration on WM-266-4 metastatic melanoma cells, which are often used in melanoma research. Due to resistance of melanoma cells high concentrations of cytotoxic substances soluble in DMSO are used in vitro tests. Consequently, total DMSO concentration often exceeds 1%. The aim of our study was to test the metabolic activity and morphology of WM-266-4 cells exposed to selected DMSO concentrations for different period of time.Cells were incubated in selected ethanol concentrations for comparison. MTT test was performed to determine proliferation activity of the cells and morphological analysis was carried out by phase-contrast microscopy. Our results show inhibitory effect of DMSO on WM-266-4 cells' metabolic activity. Morphology of the cells changed progressively with the time of exposure. Ethanol showed little effect on metabolic activity of the cells and no effect on cell morphology after selected period of time. According to our study, for specific in vitro tests concentrations of DMSO up to 1.5% may be appropriate for WM-266-4 cell line experiments.

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“…In the current study, cell viability was evaluated using the CCK-8 assay. This assay is based on dehydrogenase activity and involves a majority of high-sensitivity dehydrogenases found in cells and no significant differences in cell viability were noted among the groups at the same time points ( 41 ).…”

Section: Discussionmentioning

confidence: 99%

Osteogenic potential of cell spheroids composed of varying ratios of gingiva‑derived and bone marrow stem cells using concave microwells

Tae

Lee

et al. 2018

Exp Ther Med

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The aim of the current study was to evaluate cell viability and osteogenic differentiation potential in cell spheroids composed of varying ratios of gingiva-derived and bone marrow stem cells cultured in concave microwells. Cell spheroids were established from bone marrow and gingiva-derived stem cells in ratios of 6:0 (Group 1), 2:1 (Group 2), 3:3 (Group 3), 1:2 (Group 4), and 0:6 (Group 5). On days 3 and 5, the viability of the cell spheroids was qualitatively analyzed using a calcein acetoxymethyl ester working solution and an ethidium homodimer-1 live/dead assay. On days 1, 3, 5 and 7, a quantitative cell viability analysis was performed using a Cell Counting Kit-8. Alkaline phosphatase activity assays were performed using a commercially available kit on day 7 to assess osteogenic differentiation. In addition, reverse transcription-quantitative polymerase chain reaction and western blot analysis were performed to evaluate runt-related transcription factor 2 (Runx2) and osteocalcin expression. The ratio of gingiva-derived to bone marrow stem cells did not affect the stem cell spheroid morphology. No significant changes in cell viability were noted among the different groups following incubation for 7 days. A consistent alkaline phosphatase activity was measured in co-cultured gingiva-derived and bone marrow stem cell spheroids of varying compositions. Runx2 and osteocalcin expression was increased when co-cultured compared with pure gingiva-derived or bone marrow stem cells. In conclusion, stem cell spheroids established by co-culturing maintained morphology, viability and a high osteogenic differentiation potential during the experimental period of 7 days. These spheroids containing human gingiva-derived and bone marrow stem cells may enhance the osteogenic differentiation potential. The use of multicell spheroids may be a simple and effective strategy for improving stem cell therapy.

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

Cited by 19 publications

References 17 publications

Reactive Oxygen Species Imaging in U937 Cells

Reactive Oxygen Species Imaging in U937 Cells

The influence of dimethyl sulfoxide (DMSO) on metabolic activity and morphology of melanoma cell line WM-266-4

Osteogenic potential of cell spheroids composed of varying ratios of gingiva‑derived and bone marrow stem cells using concave microwells

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