Sensitivity of Hepatocyte Mitochondria Transmembrane Potential to Regulation with Phenylephrine after Exposure in Dimethyl Sulfoxide and 1,2-Propanediol Solutions, and Freeze-Thawing

Authors

  • Margarita Yu. Malyukina Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov
  • Nataliya S. Kavok Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov
  • Igor A. Borovoy Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov
  • Katerina A. Averchenko Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov

Keywords:

freeze-thawing, hepatocytes, dimethyl sulfoxide, 1, 2-propanediol, JC-1 probe, mitochondria

Abstract

Intensity of JC-1 probe aggregates fluorescence and agonist-induced changes in mitochondria transmembrane potential of rat hepatocytes prior to and after freeze-thawing were assessed by single-cell microfluorimetry. It was shown that incubation of the cells in 10% dimethyl sulfoxide (DMSO) and 1,2-propanediol (1,2-PD) solutions within an hour and the following washing decreased the fluorescence of dye aggregates. After freeze-thawing of cells with the cryoprotectants (2-step freezing down to the temperature of liquid nitrogen, thawing and washing-out the cryoprotectants) the intensity of dye aggregates fluorescence decreased down to 83 ± 6% of control in case of DMSO, and in the case of 1,2-PD the index made less than 10%. Unlike the fresh cell suspension where the rise of aggregates fluorescence intensity by 30% was found in response to short-term stimulation with phenylephrine (10-5 M) the hepatocyte suspension incubated in DMSO solution showed no response. After freeze-thawing of cells we revealed a partial recovery of their sensitivity to the hormone effect, but the response was recorded only after increasing the hormone concentration up to 10-4 M. Probl Cryobiol Cryomed 2013; 23(2):143-151.

Author Biographies

Margarita Yu. Malyukina, Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov

Department of Nanocrystal Matherials

Nataliya S. Kavok, Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov

Department of Nanocrystal Matherials

Igor A. Borovoy, Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov

Department of Nanocrystal Matherials

Katerina A. Averchenko, Institute for Scintillation Materials of the National Academy of Sciences of Ukraine, Kharkov

Department of Nanocrystal Matherials

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Published

2012-06-20

How to Cite

Malyukina, M. Y., Kavok, N. S., Borovoy, I. A., & Averchenko, K. A. (2012). Sensitivity of Hepatocyte Mitochondria Transmembrane Potential to Regulation with Phenylephrine after Exposure in Dimethyl Sulfoxide and 1,2-Propanediol Solutions, and Freeze-Thawing. Problems of Cryobiology and Cryomedicine, 22(2), 143–151. Retrieved from http://journal.cryo.org.ua/index.php/probl-cryobiol-cryomed/article/view/31

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Section

Theoretical and Experimental Cryobiology