Effect of dimethyl sulfoxide on cytosolic ionized calcium concentration and cytoskeletal organization of hepatocytes in a primary culture.
The addition of dimethyl sulfoxide (DMSO) to a chemically defined, serum free medium prolonged hepatocytes survival in primary culture. DMSO exposure had a remarkable effect on morphological change and F-actin filaments distribution of hepatocytes. When hepatocytes were cultured in a medium containing 2% DMSO, the cells showed a compact and cubical shape and intracellular F-actin filaments were mainly observed in a ring-like fashion around the intercellular space. After exposure to DMSO, fibronectin fibers in the interspace between cell and substratum were not apparent. Exposing the hepatocytes to DMSO also caused a sharp increase in cytosolic free ionized calcium ([Ca2+]). The initial increase in [Ca2+]i following the addition of DMSO was not attenuated by the chelation of extracellular Ca2+ with EGTA. The Ca2+ signal in the absence of extracellular Ca2+ was transient and returned to the basal levels within 1-2 min, while it was maintained at a high steady state in the presence of extracellular Ca2+. These results suggest that DMSO may be able to increase [Ca2+]i by two mechanisms, by the release of the ion from intracellular pools and, by the stimulation of influx across the plasma membrane. The increase in [Ca2+]i induced by DMSO treatment may play a role in prolonging hepatocyte survival in culture, since [Ca2+]i is one of the most important dynamic second messengers in various cellular metabolic processes.
Yamamoto
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Department of Functional Morphology, Kitasato University School of Nursing, Kanagawa, Japan.
D000818:Animals; D001572:Benzofurans; D002118:Calcium / Q000032:analysis*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects*; D003600:Cytosol / Q000032:analysis; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005353:Fibronectins / Q000378:metabolism; D016257:Fura-2; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents; D008099:Liver / Q000032:analysis / Q000166:cytology / Q000187:drug effects*; D051381:Rats; D013050:Spectrometry, Fluorescence
D001572:Benzofurans; D005353:Fibronectins; D007202:Indicators and Reagents; D002118:Calcium; D016257:Fura-2; D004121:Dimethyl Sulfoxide