Abstract
The mechanical features of individual animal cells have been regarded as indicators of cell type and state. Previously, we investigated the surface mechanics of cancer and normal stromal cells in adherent and suspended states using atomic force microscopy. Cancer cells possessed specific mechanical and actin cytoskeleton features that were distinct from normal stromal cells in adherent and suspended states. In this paper, we report the unique mechanical and actin cytoskeletal features of human embryonic kidney HEK293 cells. Unlike normal stromal and cancer cells, the surface stiffness of adherent HEK293 cells was very low, but increased after cell detachment from the culture surface. Induced actin filament depolymerization revealed that the actin cytoskeleton was the underlying source of the stiffness in suspended HEK293 cells. The exclusive mechanical response of HEK293 cells to perturbation of the actin cytoskeleton resembled that of adherent cancer cells and suspended normal stromal cells. Thus, with respect to their special cell-surface mechanical features, HEK293 cells could be categorized into a new class distinct from normal stromal and cancer cells.
Highlights
Investigation of the mechanical features provides specific information about the different states and types of animal cells
The actin cytoskeleton structures of HEK293 cells cultured under these conditions were observed by confocal laser scanning microscopy (CLSM) (Fig. 2)
Mechanical responsiveness of HEK293 cells to perturbations of the actin cytoskeleton To evaluate the contribution of F-actin structures to the mechanical properties of HEK293 cells, we examined their responsiveness to the actin cytoskeleton-modifying agents Y27632 and calyculin A
Summary
Investigation of the mechanical features provides specific information about the different states and types of animal cells. Malignant cancer cells exhibit lower stiffness than normal cells (Cross et al, 2007; Guck et al, 2005; Haghparast et al, 2013; Suresh et al, 2005); cortical stiffness changes during mitotic cell rounding (Kunda et al, 2008; Shimizu et al, 2012a); and the stiffness of the retinal epithelium changes during optic-cup morphogenesis (Eiraku et al, 2011) Such mechanical features and alterations are largely attributable to the actin cytoskeleton (Dai & Sheetz, 1995; Sugitate et al, 2009; Trickey, Vail & Guilak, 2004; Wang, 1998). It is possible to characterize the sub-membrane actin networks in each cell type as indicators of surface stiffness
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