Fukaura Eri, Kiriaki Kento, Kayier Mutailipu, Fujiwara Masashi, Takagi Mutsumi
Division of Chemistry, Graduate School of Engineering, Hokkaido University, N13W8, Kita-ku, Sapporo, 060-8628 Japan.
Cytotechnology. 2022 Feb;74(1):193-200. doi: 10.1007/s10616-021-00505-z. Epub 2021 Nov 20.
With the aim of regulating clock gene expression to control cell activities in cell processing engineering, the effect of the combination of residual glucose concentration and subsequent increment by temporal glucose feeding on the oscillation of the expression of clock gene was investigated employing rat Mesenchymal stem cell (MSC)-like cells having promoter gene with a destabilized luciferase gene (). Two experiments with several initial glucose concentrations and different times of cultures (2 and 5 days) before temporal glucose feeding (0.9 g/L) were employed to realize various concentrations of residual glucose in the medium before the feeding. In these experiments, the lower residual glucose concentrations (0.002-0.02 g/L) before temporal glucose feeding tended to induce the larger amplitude of oscillation of expression than the higher ones (0.55-0.74 g/L). When the residual glucose concentration before glucose feeding was low (0.014-0.038 g/L), the higher temporal glucose concentration (0.23-0.9 g/L) feeding tended to induce the larger amplitude of oscillation of expression than the lower ones (0.012-0.023 g/L). Taken together, we found that the amplitude of oscillation of the expression of clock gene could be controlled by the combination of residual glucose concentration and glucose concentration of subsequent temporal feeding.
为了在细胞加工工程中通过调节生物钟基因表达来控制细胞活动,利用具有不稳定荧光素酶基因启动子的大鼠间充质干细胞样细胞,研究了残余葡萄糖浓度与随后定时葡萄糖添加量的组合对生物钟基因表达振荡的影响。进行了两个实验,设置了几个初始葡萄糖浓度,并在定时葡萄糖添加(0.9 g/L)前设置了不同的培养时间(2天和5天),以实现添加前培养基中不同浓度的残余葡萄糖。在这些实验中,定时葡萄糖添加前较低的残余葡萄糖浓度(0.002 - 0.02 g/L)往往比高浓度(0.55 - 0.74 g/L)诱导更大的表达振荡幅度。当葡萄糖添加前的残余葡萄糖浓度较低(0.014 - 0.038 g/L)时,较高的定时葡萄糖浓度(0.23 - 0.9 g/L)添加往往比低浓度(0.012 - 0.023 g/L)诱导更大的表达振荡幅度。综上所述,我们发现生物钟基因表达的振荡幅度可以通过残余葡萄糖浓度和随后定时添加的葡萄糖浓度的组合来控制。