Correlation of NADH fluorescence lifetime and oxidative phosphorylation metabolism in the osteogenic differentiation of human mesenchymal stem cell

• Published in: Journal of biomedical optics Vol. 20; no. 1; p. 017004
• Main Authors: Guo, Han-Wen, Yu, Jia-Sin, Hsu, Shu-Han, Wei, Yau-Huei, Lee, Oscar K, Dong, Chen-Yuan, Wang, Hsing-Wen
• Format: Journal Article
• Language: English
• Published: United States Society of Photo-Optical Instrumentation Engineers 01.01.2015
• Subjects: Adenosine Triphosphate - metabolism; Biomedical materials; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cells, Cultured; Correlation; Differentiation; Fluorescence; Glycolysis; Humans; Lactic Acid - metabolism; Mesenchymal Stromal Cells - metabolism; Microscopy, Fluorescence - methods; NAD - metabolism; NADH; Oligomycins - pharmacology; Osteogenesis - drug effects; Osteogenesis - physiology; Oxidative Phosphorylation; Oxygen Consumption; Phosphorylation; Stem cells; mitochondrial function; metabolism; human mesenchymal stem cell differentiation; NADH fluorescence lifetime; oxygen consumption; adenosine triphosphate level
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.JBO.20.1.017004

Reduced nicotinamide dinucleotide (NADH) fluorescence lifetime has been broadly used as a metabolic indicator for stem cell imaging. However, the direct relationship between NADH fluorescence lifetime and metabolic pathway and activity remains to be clarified. In this study, we measured the NADH fluorescence lifetime of human mesenchymal stem cells (hMSCs) as well as the metabolic indictors, such as adenosine triphosphate (ATP) level, oxygen consumption, and lactate release, up to 4 weeks under normal osteogenic differentiation and oxidative phosphorylation-attenuated/inhibited differentiation by oligomycin A (OA) treatment. NADH fluorescence lifetime was positively correlated with oxygen consumption and ATP level during energy transformation from glycolysis to oxidative phosphorylation. Under OA treatment, oxidative phosphorylation was attenuated/inhibited (i.e., oxygen consumption remained the same as controls or lower), cells showed attenuated differentiation under glycolysis, and NADH fluorescence lifetime change was not detected. Increased expression of the overall complex proteins was observed in addition to Complex I. We suggested special caution needs to be exercised while interpreting NADH fluorescence lifetime signal in terms of stem cell differentiation.