Oxidative stress triggers cytokinesis failure in hepatocytes upon isolation

Free Radical Research
Ana M TormosJuan Sastre

Abstract

Primary hepatocytes are highly differentiated cells and proliferatively quiescent. However, the stress produced during liver digestion seems to activate cell cycle entry by proliferative/dedifferentiation programs that still remain unclear. The aim of this work was to assess whether the oxidative stress associated with hepatocyte isolation affects cell cycle and particularly cytokinesis, the final step of mitosis. Hepatocytes were isolated from C57BL/6 mice by collagenase perfusion in the absence and presence of N-acetyl cysteine (NAC). Polyploidy, cell cycle, and reactive oxygen species (ROS) were studied by flow cytometry (DNA, phospho-histone 3, and CellROX(®) Deep Red) and Western blotting (cyclins B1 and D1, and proliferating cell nuclear antigen). mRNA expression of cyclins A1, B1, B2, D1, and F by reverse transcription (RT)-PCR was also assessed. Glutathione levels were measured by mass spectrometry. Here we show that hepatocyte isolation enhanced cell cycle entry, increased hepatocyte binucleation, and caused marked glutathione oxidation. Addition of 5 mM NAC to the hepatocyte isolation media prevented glutathione depletion, partially blocked ROS production and cell cycle entry of hepatocytes, and avoided the blockade o...Continue Reading

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Citations

Mar 26, 2016·Mechanisms of Ageing and Development·Grasiella A AndrianiCristina Montagna
Jun 28, 2015·Biochemical and Biophysical Research Communications·Jaeyong LeeJunichi Fujii
Jul 8, 2016·Experimental Biology and Medicine·Katrin ZeilingerFanny Knöspel
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Aug 25, 2021·ACS Chemical Biology·Arnout G SchepersSangeeta N Bhatia

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