Quantitative metabolic imaging using endogenous fluorescence to detect stem cell differentiation

TLDR

Non‑invasive, high‑resolution mapping of cellular metabolism can accelerate stem‑cell therapy evaluation and tissue development studies, offering a rapid, high‑content alternative to destructive, time‑consuming traditional assays. The study aims to use two‑photon excited fluorescence microscopy to characterize how endogenous fluorescence and redox state correlate with human mesenchymal stem cell differentiation into adipogenic and osteoblastic lineages. By measuring the optical redox ratio FAD/(NADH+FAD) with liquid chromatography/mass spectrometry, quantitative PCR, and fluorescence imaging, the authors assess its sensitivity to metabolic shifts and link decreases in the ratio to the onset of differentiation.

Abstract

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.