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Reducing environment protects sinusoidal lymphocytes isolated from normal human liver from apoptosis
K Kinoshita1, M Kanai, M Awane
1Second Department of Surgery, College of Medical Technology, Kyoto University, Japan.
Journal of Hepatology
|January 1, 1997
Summary
Human liver sinusoidal mononuclear cells undergo apoptosis ex vivo. This cell death is reversed by reducing agents, suggesting a sensitivity to redox environment distinct from peripheral blood mononuclear cells.
Area of Science:
- Immunology
- Hepatology
- Cell Biology
Background:
- Human liver sinusoidal mononuclear cells (HSMCs) exhibit distinct lymphocyte populations and antigen expression compared to peripheral blood mononuclear cells (PBMCs).
- Previous attempts to culture HSMCs failed due to rapid cell death under standard in vitro conditions.
Purpose of the Study:
- To investigate the characteristics of HSMC cell death after isolation.
- To identify culture conditions that improve HSMC viability.
Main Methods:
- HSMCs were isolated from University of Wisconsin solution perfused from human donor livers.
- Cell viability was assessed over 48 hours under various culture conditions.
- Apoptosis was confirmed via DNA laddering and electron microscopy.
- The effects of human liver extracts, 2-mercaptoethanol, reduced glutathione, and diamide were evaluated.
Main Results:
- 70% of isolated HSMCs died within 48 hours in culture, compared to 10% of PBMCs.
- HSMC death exhibited features of apoptosis, including DNA laddering and characteristic morphology.
- Human liver extracts, 2-mercaptoethanol, and reduced glutathione significantly reduced HSMC apoptosis.
- Diamide, an oxidant, reversed the protective effect of liver extract.
Conclusions:
- HSMCs are more prone to apoptosis ex vivo than PBMCs.
- Exogenous agents that create reducing conditions can rescue HSMCs from apoptosis.
- HSMCs may possess a unique sensitivity to the redox environment compared to PBMCs.