Nanomechanobiology of human mesenchymal stem cells (hMSCs) was studied using atomic force microscopy (AFM). MSCs are progenitors of connective tissue, including bone and cartilage. Cells were differentiated into bone and cartilage type cells using specific media constituents. Typical height and deflection images of a hMSC and hMSC-Ob can be seen in the figure. Force-volume mode of AFM evaluated elasticity of hMSCs, hMSC-derived osteoblasts (hMSC-Obs) and hMSC-derived chondrocytes (hMSC-Chs) before, during and after treatment with Cytochalasin D (CytD), an inhibitor of actin filament polymerization. CytD treatment resulted in morphology changes of all cell types, with a decrease in elastic modulus of hMSCs and hMSC-Chs. The effect of CytD was less pronounced in hMSC-Obs. Removal of drug from media with cytoskeleton disrupted hMSC-Chs resulted in cell’s ability to regain cytoskeletal structure, reverting to original shape. This was confirmed using fluorescence with cytoskeleton rescue by removal of CytD, washing and replacement with non-CytD media. The use of AFM to determine nanomechanical properties of cells can be used to better define differentiating stem cells changing into mechanically distinct specialized
cells as well as long-term study of drug effects on cytoskeleton.
Xanya Sofra Weiss
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