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<p>Differentiation of the human monocytic leukemia cell line THP-1 is widely used to analyze the function of monocyte/macrophage-like cells&#xa0;<i>in vitro</i>. Although chemotaxis, a critical function of monocytes/macrophages enabling tissue accumulation, has been extensively studied, methods to evaluate sustained, long-distance chemotaxis remain underexplored. Therefore, we aimed to evaluate macrophage-like cells&#xa0;<i>in vitro&#xa0;</i>by differentiating THP-1 cells into monocyte/macrophage-like cells exhibiting sustained, strong chemotaxis over long distances (up to 260&#xa0;μm). Using various reagents, we identified the combination of vitamin D, panobinostat, and granulocyte–macrophage-colony-stimulating factor as optimal for achieving high directionality and velocity in cell migration, as analyzed using the TAXIScan cell dynamics assay device. The differentiated cells matured into M1 macrophage-like cells and displayed reduced migratory capacity post-maturation, along with enhanced phagocytosis and reactive oxygen species production. Collectively, our differentiation and analysis methods provide a reliable platform for basic research into cellular maturation processes and drug development targeting the regulation of monocyte/macrophage dynamics.</p>

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Establishment of a method to evaluate the dynamics of highly chemotactic THP-1 cells during differentiation into monocyte-M1 macrophage-like cells

  • 期刊论文

  • 发表日期：

    2025年7月30日

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  • Shuichiro Okamoto,
  • Kei Miyano,
  • Yasumitsu Nishimura,
  • Nahoko Tomonobu,
  • Rie Kinoshita,
  • Masakiyo Sakaguchi,
  • Einosuke Ikeshita,
  • Ayasa Kamezaki,
  • Aya Morihara,
  • Futoshi Kuribayashi,
  • Akira Yamauchi

摘要

Differentiation of the human monocytic leukemia cell line THP-1 is widely used to analyze the function of monocyte/macrophage-like cells in vitro. Although chemotaxis, a critical function of monocytes/macrophages enabling tissue accumulation, has been extensively studied, methods to evaluate sustained, long-distance chemotaxis remain underexplored. Therefore, we aimed to evaluate macrophage-like cells in vitro by differentiating THP-1 cells into monocyte/macrophage-like cells exhibiting sustained, strong chemotaxis over long distances (up to 260 μm). Using various reagents, we identified the combination of vitamin D, panobinostat, and granulocyte–macrophage-colony-stimulating factor as optimal for achieving high directionality and velocity in cell migration, as analyzed using the TAXIScan cell dynamics assay device. The differentiated cells matured into M1 macrophage-like cells and displayed reduced migratory capacity post-maturation, along with enhanced phagocytosis and reactive oxygen species production. Collectively, our differentiation and analysis methods provide a reliable platform for basic research into cellular maturation processes and drug development targeting the regulation of monocyte/macrophage dynamics.