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<p>This study aimed to develop a simplified microfluidic culture system for HepG2 cells and to investigate the effects of fluid shear stress (FSS) on cellular morphology, proliferation, nuclear morphology, and autophagy-related responses. A PDMS-based microfluidic device was fabricated using soft lithography and integrated into a closed-loop perfusion system. HepG2 cells were cultured under static or flow conditions, with continuous medium perfusion at a defined flow velocity of 2.42 mm/s for 24 h to generate low FSS. A streamlined microfluidic platform was successfully fabricated and applied to generate controlled dynamic culture conditions. Continuous perfusion at a flow velocity of 2.42 mm/s for 24 h produced low FSS ranging from 0.014 to 0.04 dyn/cm². Under flow conditions, HepG2 cell numbers were significantly reduced compared with those in static culture. Morphologically, cells exposed to FSS exhibited a transition from polygonal to more elongated shapes. Nuclear area increased by approximately 1.4-fold under flow conditions. In addition, immunofluorescence analysis revealed a marked upregulation of the autophagy-related gene ATG5 in response to FSS. The proposed microfluidic system provides a simple platform for investigating the effects of low FSS on HepG2 cells.</p> Graphical abstract <p></p>

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Introduction to the simple setup microfluidic system for the dynamic culture of HepG2 cell line

  • 期刊论文

  • 发表日期：

    2026年4月7日

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  • Hoang Mai Thanh Truc,
  • Huynh Huu Tinh,
  • Truong Hai Nhung,
  • Thanh-Qua Nguyen,
  • Le Van Trinh

摘要

This study aimed to develop a simplified microfluidic culture system for HepG2 cells and to investigate the effects of fluid shear stress (FSS) on cellular morphology, proliferation, nuclear morphology, and autophagy-related responses. A PDMS-based microfluidic device was fabricated using soft lithography and integrated into a closed-loop perfusion system. HepG2 cells were cultured under static or flow conditions, with continuous medium perfusion at a defined flow velocity of 2.42 mm/s for 24 h to generate low FSS. A streamlined microfluidic platform was successfully fabricated and applied to generate controlled dynamic culture conditions. Continuous perfusion at a flow velocity of 2.42 mm/s for 24 h produced low FSS ranging from 0.014 to 0.04 dyn/cm². Under flow conditions, HepG2 cell numbers were significantly reduced compared with those in static culture. Morphologically, cells exposed to FSS exhibited a transition from polygonal to more elongated shapes. Nuclear area increased by approximately 1.4-fold under flow conditions. In addition, immunofluorescence analysis revealed a marked upregulation of the autophagy-related gene ATG5 in response to FSS. The proposed microfluidic system provides a simple platform for investigating the effects of low FSS on HepG2 cells.

Graphical abstract