Decreased cell viability resulting from a severe condition of nutrients deprivation and hypoxia has been the major obstacle in three-dimensional (3D) tissue construction. Therefore, technical improvement which prevents cell death caused by starvation and low oxygen is desired for the development of large, thick tissues. We focused on the anti-glycolytic effect of resveratrol (RSV), a naturally-occurring polyphenol known as a caloric restriction mimetic, and investigated its cytoprotective effect in two-dimensional (2D) and 3D-cell culture using H9c2 rat myoblast cells. Glucose deprivation by culturing with low glucose media caused time- and dose-dependent cell death in H9c2 cells. In contrast, RSV treatment at 100 μM significantly increased the cell viability by preventing cell death. RSV showed anti-glycolytic effect associated with a down-regulation of glycolytic genes (GLUT1, PKM2) and glucose uptake activity, and increased the activation of AMP-activated protein kinase (AMPK), an essential cellular energy sensor activated in the condition of energy deprivation. RSV treatment markedly improved the viability of myoblast cells cultured in a hypoxic, low glucose condition and attenuated the up-regulation of glycolytic genes by hypoxic response. In 3D-cultured model, spheroids constructed with RSV-treated cells showed improved cell viability and intact histological appearance compared with control. These results suggest that glycolytic inhibition by RSV decreases the glucose usage of myoblast cells, therefore, prevents cell death caused by nutrient deprivation and hypoxic condition. Our finding provides useful information to improve cell viability in a condition that nutrients and oxygen are low in supply, and be a possible application to the 3D-tissue construction.

由于营养缺乏和缺氧的严重状况而导致的细胞活力下降已成为三维（3D）组织构建的主要障碍。因此，对于大而厚的组织的发育，需要防止由饥饿和低氧引起的细胞死亡的技术改进。我们专注于白藜芦醇（RSV）（一种天然的多酚，称为热量限制模拟物）的抗糖酵解作用，并研究了其在使用H9c2大鼠成肌细胞的二维（2D）和3D细胞培养中的细胞保护作用。通过在低葡萄糖培养基中培养而剥夺葡萄糖会导致H9c2细胞中时间和剂量依赖性细胞死亡。相反，以100μM的RSV处理可通过防止细胞死亡来显着提高细胞活力。RSV显示出与糖酵解基因（GLUT1，PKM2）下调和葡萄糖摄取活性相关的抗糖酵解作用，并增加了AMP激活的蛋白激酶（AMPK）的激活，这是一种在能量条件下被激活的重要细胞能量传感器。剥夺。RSV处理显着提高了在低氧，低葡萄糖条件下培养的成肌细胞的活力，并通过低氧反应减弱了糖酵解基因的上调。在3D培养的模型中，与对照组相比，用RSV处理的细胞构建的球体显示出更高的细胞活力和完整的组织学外观。这些结果表明，RSV的糖酵解抑制作用降低了成肌细胞的葡萄糖使用量，因此，可以防止因营养剥夺和低氧状态导致的细胞死亡。