A microfluidic cell culture system for monitoring of sequential changes in endothelial cells after heat stress

Abstract

Endothelial damage induced by a highly elevated body temperature is crucial in some diseases including viral hemorrhagic fevers. Here, we report the heat-induced sequential changes of endothelial cells under shear stress, which were determined with a microfluidic culture system. Although live cell imaging showed only minor changes in the appearance of heat-treated cells, Hsp70 mRNA expression analysis demonstrated that the endothelial cells in channels of the system responded well to heat treatment. F-actin staining also revealed clear changes in the bundles of actin filaments after heat treatment. Well-organized bundles of actin filaments in control cells disappeared in heat-treated cells cultured in the channel. Furthermore, the system enabled detection of sequential changes in plasminogen activator inhibitor-1 (PAI-1) secretion from endothelial cells. PAI-1 concentration in the effluent solution was significantly elevated for the first 15min after initiation of heat treatment, and then decreased subsequently. This study provides fundamental information on heat-induced endothelial changes under shear stress and introduces a potent tool for analyzing endothelial secretions.

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“…40 In particular, heat-stressed endothelial cells showed increased Hsp70 mRNA expression, which is mainly involved in partial refolding of damaged proteins, protein disposal and aggregation prevention. 41 Higher temperatures could lead to Hsp70 exhaustion and accumulation of damaged proteins. Studies on magnetic nanoparticles already showed their high heating capacity upon application of alternating magnetic field.…”

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“…8 The microchip cell culture system enables continuous and rigorous flow control, which is advantageous in vascular biology studies. [9][10][11] Vascular dysfunction 12 is associated with major diseases such as diabetes and cancer, and microfluidic cell culture systems are excellent tools for elucidating the cellular mechanisms underlying these diseases. A microfluidic cell culture system has been developed as a perfusion system that mimics the in-vivo environment 13,14 and facilitates evaluation of shear stress.…”