Direct monitoring of drug-induced mechanical response of individual cells by atomic force microscopy

Abstract

Mechanical characteristics of individual cells play a vital role in many biological processes and are considered as indicators of the cells' states. Disturbances including methyl-β-cyclodextrin (MβCD) and cytochalasin D (cytoD) are known to significantly affect the state of cells, but little is known about the real-time response of single cells to these drugs in their physiological condition. Here, nanoindentation-based atomic force microscopy (AFM) was used to measure the elasticity of human embryonic kidney cells in the presence and absence of these pharmaceuticals. The results showed that depletion of cholesterol in the plasma membrane with MβCD resulted in cell stiffening whereas depolymerization of the actin cytoskeleton by cytoD resulted in cell softening. Using AFM for real-time measurements, we observed that cells mechanically responded right after these drugs were added. In more detail, the cell´s elasticity suddenly increased with increasing instability upon cholesterol extraction while it is rapidly decreased without changing cellular stability upon depolymerizing actin cytoskeleton. These results demonstrated that actin cytoskeleton and cholesterol contributed differently to the cell mechanical characteristics.

Keywords: HEK cells; atomic force microscopy; cell elasticity; nanoindentation; stiffness.