Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs

Abstract

Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed to identify the biological and physical factors involved in the differences in freeze–thaw tolerance between MSCs and HUVECs. Materials and methods For biological analysis, MSC and HUVEC viability after freeze-thawing and alteration of gene expression in response to dimethyl sulfoxide (DMSO, a cryoprotectant) were quantitatively evaluated. For physical analysis, the cell membrane fluidity of MSCs and HUVECs before and after DMSO addition was assessed using a histogram for generalized polarization frequency. Results HUVECs showed lower live cell rates and higher gene expression alteration related to extracellular vesicles in response to DMSO than MSCs. Fluidity measurements revealed that the HUVEC membrane was highly fluidic and sensitive to DMSO compared to that of MSCs. Addition of CAY10566, an inhibitor of stearoyl-coA desaturase (SCD1) that produces highly fluidic desaturated fatty acids, decreased the fluidity of HUVECs and increased their tolerance to DMSO. The combination of CAY10566 and antioxidant glutathione (GSH) treatment improved HUVEC viability from 57 to 69%. Membrane fluidity alteration may thus contribute to pore-induced DMSO influx into the cytoplasm and reactive oxygen species production, leading to greater cytotoxicity in HUVECs, which have low antioxidant capacity. Conclusions Differences in freeze–thaw tolerance originate from differences in the cell membranes with respect to fluidity and antioxidant capacity. These findings provide a basis for analyzing cell biology and membrane-physics to establish appropriate long-term preservation methods aimed at promoting transplantation therapies.

Vascular Smooth Muscle TRPV4 (Transient Receptor Potential Vanilloid Family Member 4) Channels Regulate Vasoconstriction and Blood Pressure in Obesity

“…We found a higher EC 50 of GSK101 in SMCs than ECs, suggesting different sensitivity of TRPV4 to its agonist GSK101 in different cell types. 32,33 In the present study, ECs seems more sensitive to GSK101 (EC 50 ≈2.28 nmol/L) than SMCs (EC 50 ≈43.49 nmol/L). It is likely that one type of ion channel can show different sensitivity to its activator in different cell types.…”

Mechanisms of Arachidonic Acid In Vitro Tumoricidal Impact

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Cytotoxic Activity of Herbal Medicines as Assessed in Vitro: A Review

“…Numerous organic solvents, without toxic effects on cells, are widely used for dissolving the hydrophobic compounds in different types of cell culture assays [79–81] . Therefore, in a preliminary assay, it is important to determine the maximum or minimum tolerable concentration for solvent on cell toxicity that can be used to prepare solutions of herbal extracts or other compounds [82–85] . Dimethyl sulfoxide (DMSO), ethanol and methanol are suitable solvents widely used as vehicles for drug delivery, [86] crude extraction, and dissolving various plant extracts for enhanced solubility in biological studies [87] .…”