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Quantitative metabolic imaging using endogenous fluorescence to detect stem cell differentiation

机译：使用内源性荧光定量代谢成像检测干细胞分化

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The non-invasive high-resolution spatial mapping of cell metabolism within tissues could provide substantial advancements in assessing the efficacy of stem cell therapy and understanding tissue development. Here, using two-photon excited fluorescence microscopy, we elucidate the relationships among endogenous cell fluorescence, cell redox state, and the differentiation of human mesenchymal stem cells into adipogenic and osteoblastic lineages. Using liquid chromatography/mass spectrometry and quantitative PCR, we evaluate the sensitivity of an optical redox ratio of FAD/(NADH + FAD) to metabolic changes associated with stem cell differentiation. Furthermore, we probe the underlying physiological mechanisms, which relate a decrease in the redox ratio to the onset of differentiation. Because traditional assessments of stem cells and engineered tissues are destructive, time consuming, and logistically intensive, the development and validation of a non-invasive, label-free approach to defining the spatiotemporal patterns of cell differentiation can offer a powerful tool for rapid, high-content characterization of cell and tissue cultures.
机译：组织内细胞代谢的非侵入性高分辨率空间作图可以在评估干细胞疗法的疗效和了解组织发育方面提供重大进展。在这里，使用双光子激发荧光显微镜，我们阐明了内源性细胞荧光，细胞氧化还原状态和人间充质干细胞向成脂和成骨细胞谱系分化之间的关系。使用液相色谱/质谱和定量PCR，我们评估FAD /（NADH + FAD）的光学氧化还原比对与干细胞分化相关的代谢变化的敏感性。此外，我们探讨了潜在的生理机制，这些机制将氧化还原比的降低与分化的开始联系起来。由于对干细胞和工程组织的传统评估是破坏性的，耗时的和后勤性的，因此开发和验证无创，无标签的方法来定义细胞分化的时空模式可以为快速，高效率地提供强大的工具。细胞和组织培养物的含量表征。

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