Mechanical properties of human amniotic fluid stem cells using nanoindentation.

The aim of this study was to obtain nanomechanical properties of living cells focusing on human amniotic fluid stem (hAFS) cell using nanoindentation techniques. We modified the conventional method of atomic force microscopy (AFM) in aqueous environment for cell imaging and indentation to avoid inherent difficulties. Moreover, we determined the elastic modulus of murine osteoblast (OB6) cells and hAFS cells at the nucleus and cytoskeleton using force-displacement curves and Hertz theory. Since OB6 cell line has been widely used, it was selected to validate and compare the obtained results with the previous research studies. As a result, we were able to capture high resolution images through utilization of the tapping mode without adding protein or using fixation methods. The maximum depth of indentation was kept below 15% of the cell thickness to minimize the effect of substrate hardness. Nanostructural details on the surface of cells were visualized by AFM and fluorescence microscopy. The cytoskeletal fibers presented remarkable increase in elastic modulus as compared with the nucleus. Furthermore, our results showed that the elastic modulus of hAFS cell edge (31.6 kPa) was lower than that of OB6 cell edge (42.2 kPa). In addition, the elastic modulus of nucleus was 13.9 kPa for hAFS cell and 26.9 kPa for OB6 cells. Differences in cell elastic modulus possibly resulted from the type and number of actin cytoskeleton organization in these two cell types.