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Molecular Beacon Imaging System to Discriminate the Differentiation State of Cells from Energy Metabolic Pathways
ACS Sensors ( IF 8.2 ) Pub Date : 2023-05-30 , DOI: 10.1021/acssensors.3c00106 Yuki Murata 1 , Jun-Ichiro Jo 1 , Yasuhiko Tabata 1
ACS Sensors ( IF 8.2 ) Pub Date : 2023-05-30 , DOI: 10.1021/acssensors.3c00106 Yuki Murata 1 , Jun-Ichiro Jo 1 , Yasuhiko Tabata 1
Affiliation
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Metabolic pathways of energy production play an essential role as a function of cells. It is well recognized that the differentiation state of stem cells is highly associated with their metabolic profile. Therefore, visualization of the energy metabolic pathway makes it possible to discriminate the differentiation state of cells and predict the cell potential for reprogramming and differentiation. However, at present, it is technically difficult to directly assess the metabolic profile of individual living cells. In this study, we developed an imaging system of cationized gelatin nanospheres (cGNS) incorporating molecular beacons (MB) (cGNSMB) to detect intracellular pyruvate dehydrogenase kinase 1 (PDK1) and peroxisome proliferator-activated receptor γ, coactivator-1α (PGC-1α) mRNA of key regulators in the energy metabolism. The prepared cGNSMB was readily internalized into mouse embryonic stem cells, while their pluripotency was maintained. The high level of glycolysis in the undifferentiated state, the increased oxidative phosphorylation over the spontaneous early differentiation, and the lineage-specific neural differentiation were visualized based on the MB fluorescence. The fluorescence intensity corresponded well to the change of extracellular acidification rate and the oxygen consumption rate of representative metabolic indicators. These findings indicate that the cGNSMB imaging system is a promising tool to visually discriminate the differentiation state of cells from energy metabolic pathways.
中文翻译：
分子信标成像系统从能量代谢途径辨别细胞的分化状态
能量产生的代谢途径作为细胞的功能发挥着重要作用。众所周知，干细胞的分化状态与其代谢特征高度相关。因此，能量代谢途径的可视化使得区分细胞的分化状态并预测细胞重编程和分化的潜力成为可能。然而，目前，直接评估单个活细胞的代谢特征在技术上还很困难。在这项研究中，我们开发了一种结合分子信标 (MB) 的阳离子明胶纳米球 (cGNS) 成像系统 (cGNS MB )，用于检测细胞内丙酮酸脱氢酶激酶 1 (PDK1) 和过氧化物酶体增殖物激活受体 γ、辅激活剂-1α (PGC- 1α) 能量代谢关键调节因子的mRNA。制备的 cGNS MB很容易内化到小鼠胚胎干细胞中，同时保持其多能性。基于MB荧光可视化未分化状态下的高水平糖酵解、自发早期分化中氧化磷酸化的增加以及谱系特异性神经分化。荧光强度与细胞外酸化率和代表性代谢指标耗氧率的变化具有良好的对应关系。这些发现表明，cGNS MB成像系统是一种很有前途的工具，可以从视觉上区分细胞的分化状态与能量代谢途径。
更新日期：2023-05-30
中文翻译：
分子信标成像系统从能量代谢途径辨别细胞的分化状态
能量产生的代谢途径作为细胞的功能发挥着重要作用。众所周知，干细胞的分化状态与其代谢特征高度相关。因此，能量代谢途径的可视化使得区分细胞的分化状态并预测细胞重编程和分化的潜力成为可能。然而，目前，直接评估单个活细胞的代谢特征在技术上还很困难。在这项研究中，我们开发了一种结合分子信标 (MB) 的阳离子明胶纳米球 (cGNS) 成像系统 (cGNS MB )，用于检测细胞内丙酮酸脱氢酶激酶 1 (PDK1) 和过氧化物酶体增殖物激活受体 γ、辅激活剂-1α (PGC- 1α) 能量代谢关键调节因子的mRNA。制备的 cGNS MB很容易内化到小鼠胚胎干细胞中，同时保持其多能性。基于MB荧光可视化未分化状态下的高水平糖酵解、自发早期分化中氧化磷酸化的增加以及谱系特异性神经分化。荧光强度与细胞外酸化率和代表性代谢指标耗氧率的变化具有良好的对应关系。这些发现表明，cGNS MB成像系统是一种很有前途的工具，可以从视觉上区分细胞的分化状态与能量代谢途径。
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