Mechanical Systems and Engineering Graduate School Systems Design, Tokyo Metropolitan University, 1-1 Minamiosawa, Hachioji, Tokyo, 192-0397, Japan.
Laboratory of Biomedical and Biophysical Chemistry, Institute for Materials Chemistry and Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka, 819-0395, Japan.
Anal Sci. 2021;37(3):447-451. doi: 10.2116/analsci.20SCP02.
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.
为了制定细胞培养基质设计的指导方针以控制人类间充质干细胞（MSC）分化，我们定量表征了 MSC 对培养基质弹性模量的响应异质性。我们使用具有可控弹性模量的苯乙烯化明胶凝胶分析了三个不同 MSC 批次中机械转导蛋白 YAP 和成骨分化因子 RUNX2 对弹性模量的依赖性动力学。核内具有 YAP 的细胞百分比随弹性模量的增加呈线性增加，对于所有分析的批次均在 10 kPa 处达到平台期。随着基质弹性模量的增加，百分比呈相同的线性函数增加。具有 RUNX2 核定位的细胞百分比也随基质弹性模量的增加呈线性增加，在 5 kPa 处达到平台期，尽管批次之间的线性增加区域的回归线不同。细胞群体中 YAP 和 RUNX2 动力学的这些相似性和差异是设计基质弹性模量以操纵 YAP 和 RUNX2 定位的基础。
