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Reactive Oxygen Species Imaging in U937 Cells
Ankush Prasad1, Michaela Sedlářová2, Anastasiia Balukova1
1Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czechia.
Abstract:
The U937 cell culture is a pro-monocytic, human histiocytic lymphoma cell line. These monocytes can differentiate into either macrophages or dendritic cells (antigen-presenting cells) depending on the initiators. The U937 cells activated in the presence of phorbol 12-myristate 13-acetate (PMA) change their morphology into macrophage-like cells creating pseudopodia and adhering generously. Macrophages are known to produce reactive oxygen species (ROS) mostly during phagocytosis of foreign particles, an important non-specific immune response. Recently, we have focused on the role of hydroxyl radical (HO∙) and provide evidence on its importance for differentiation in U937 cells. Based on electron paramagnetic resonance (EPR) spectroscopy combined with confocal laser scanning microscopy (CLSM), formation of HO∙ was confirmed within the cells undergoing differentiation and/or apoptosis during the PMA treatment. This study aims to increase our knowledge of ROS metabolism in model cell lines used in human research.
Insights
Hydroxyl radical (HO∙) plays a key role in U937 cell differentiation into macrophage-like cells. This study confirms HO∙ formation during phorbol 12-myristate 13-acetate (PMA) induced differentiation using advanced microscopy and spectroscopy.
Area of Science:
- Cell Biology
- Immunology
- Biochemistry
Background:
- U937 cells are a human histiocytic lymphoma cell line with pro-monocytic properties.
- These cells can differentiate into macrophages or dendritic cells, crucial for immune responses.
- Reactive oxygen species (ROS), including hydroxyl radical (HO∙), are implicated in cellular processes.
Purpose of the Study:
- To investigate the role of hydroxyl radical (HO∙) in U937 cell differentiation.
- To elucidate the contribution of ROS metabolism to U937 cell differentiation processes.
- To enhance understanding of ROS in model cell lines for human research.
Main Methods:
- U937 cell culture and differentiation induction using phorbol 12-myristate 13-acetate (PMA).
- Electron paramagnetic resonance (EPR) spectroscopy to detect radical formation.
- Confocal laser scanning microscopy (CLSM) for cellular visualization.
Main Results:
- Phorbol 12-myristate 13-acetate (PMA) treatment induced macrophage-like morphology in U937 cells.
- Formation of hydroxyl radical (HO∙) was confirmed within differentiating U937 cells.
- HO∙ generation was observed during both differentiation and apoptosis phases.
Conclusions:
- Hydroxyl radical (HO∙) is a significant factor in U937 cell differentiation.
- ROS metabolism, particularly HO∙, is integral to U937 cell differentiation.
- This research contributes to understanding ROS roles in immune cell differentiation models.
