Designing Elastic Modulus of Cell Culture Substrate to Regulate YAP and RUNX2 Localization for Controlling Differentiation of Human Mesenchymal Stem Cells

Abstract

To establish a guideline for the design of cell culture substrates to control human mesenchymal stem cell (MSC) differentiation, we quantitatively characterized the heterogeneity in the responsiveness of MSCs to the elastic modulus of culture substrates. We analyzed the elastic modulus-dependent dynamics of a mechanotransducer, YAP, and an osteogenic differentiation factor, RUNX2, in three different MSC lots using a styrenated gelatin gel with controllable elastic modulus. The percentage of cells with YAP in the nucleus increased linearly with increases in the elastic modulus, reaching a plateau at 10 kPa for all the lots analyzed. The increase in the percentage with the substrate elastic modulus was described by the same linear function. The percentage of cells with RUNX2 nuclear localization also increased linearly with increases in the substrate elastic modulus, plateauing at 5 kPa, although the regression lines to the linearly increasing regions varied between lots. These similarities and differences in YAP and RUNX2 dynamics among cell populations are basis to design the substrate elastic modulus to manipulate YAP and RUNX2 localizations.

Silk fibroin films with embedded magnetic nanoparticles: evaluation of the magneto-mechanical stimulation effect on osteogenic differentiation of stem cells

“…62,63 Above all, the substrate stiffness is a potent regulator of the differentiation process of stem cells. 64–66…”

“…62,63 Above all, the substrate stiffness is a potent regulator of the differentiation process of stem cells. [64][65][66] A clear influence of the applied magnetic field on osteogenic cellular differentiation is revealed only by the analysis of the gene expression markers (Fig. 8).…”

Optimization of Mechanosensitive Cross-Talk between Matrix Stiffness and Protein Density: Independent Matrix Properties Regulate Spreading Dynamics of Myocytes

“…YAP is a transcriptional regulator that requires Rho activity to regulate mechanical properties of the ECM and cell geometry [45]. In mesenchymal stem cells, the percentage of cells with YAP in the nucleus increases linearly with increases in the elastic modulus, reaching a plateau at 10 kPa [46]. Considering a comparable YAP dependence in C2C12 cells, our findings suggest a potential link between cell spreading duration after proliferation with the content of YAP in the nucleus (Figure 7H), while cell spreading speed is solely regulated by the amount of matrix proteins in the ECM (Figure 7G).…”

The Oncogenic and Tumor Suppressive Long Non-Coding RNA–microRNA–Messenger RNA Regulatory Axes Identified by Analyzing Multiple Platform Omics Data from Cr(VI)-Transformed Cells and Their Implications in Lung Cancer

“…miR-182-5p can repress bladder cancer cell migration, invasion, and colony formation [ 47 ]. The GOLGA8B/miR-30d-5p/RUNX2 regulatory axis has important functions in lung cancer cell proliferation and stemness via regulatory and physical interactions of GOLGA8B and RUNX2 with both stemness-related genes and stemness TFs [ 48 ]. In addition, the oncogenic PDCD6IPP2/miR-23a-3p/HOXA1 axis shows a highly significant stemness signature due to the predicted interaction of HOXA1 with stemness-related genes and stemness TFs at physical and regulatory levels.…”