A comparative study of metabolic state of stem cells during osteogenic and adipogenic differentiations via fluorescence lifetime imaging microscopy

Sandeep Chakraborty; Meng-Hsin Ou; Jean-Cheng Kuo; Arthur Chiou

doi:10.1117/12.2246255

31 October 2016 A comparative study of metabolic state of stem cells during osteogenic and adipogenic differentiations via fluorescence lifetime imaging microscopy

Sandeep Chakraborty, Meng-Hsin Ou, Jean-Cheng Kuo, Arthur Chiou

Author Affiliations +

Proceedings Volume 10024, Optics in Health Care and Biomedical Optics VII; 100243Y (2016) https://doi.org/10.1117/12.2246255
Event: SPIE/COS Photonics Asia, 2016, Beijing, China

Abstract

Cellular metabolic state can serve as a biomarker to indicate the differentiation potential of stem cells into other specialized cell lineages. In this study, two-photon fluorescence lifetime imaging microscopy (2P-FLIM) was applied to determine the fluorescence lifetime and the amounts of the auto-fluorescent metabolic co-factor reduced nicotinamide adenine dinucleotide (NADH) to elucidate the cellular metabolism of human mesenchymal stem cells (hMSCs) in osteogenic and adipogenic differentiation processes. 2P-FLIM provides the free to protein-bound NADH ratio which can serve as the indicator of cellular metabolic state. We measured NADH fluorescence lifetime at 0, 7, and 14 days after hMSCs were induced for either osteogenesis or adipogenesis. In both cases, the average fluorescence lifetime increased significantly at day 14 (P < 0.001), while the ratio of free to protein-bound NADH ratio decreased significantly in 7- days (P < 0.001) and 14-days (P < 0.001). Thus, our results indicated a higher metabolic rate in both osteogenic and adipogenic differentiation processes when compared with undifferentiated hMSCs. This approach may be further utilized to study proliferation efficiency and differentiation potential of stem cells into other specialized cell lineages.

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Sandeep Chakraborty, Meng-Hsin Ou, Jean-Cheng Kuo, and Arthur Chiou "A comparative study of metabolic state of stem cells during osteogenic and adipogenic differentiations via fluorescence lifetime imaging microscopy", Proc. SPIE 10024, Optics in Health Care and Biomedical Optics VII, 100243Y (31 October 2016); https://doi.org/10.1117/12.2246255

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KEYWORDS

Luminescence

Stem cells

Fluorescence lifetime imaging

Mode conditioning cables

Microscopy

Data analysis

Molecules

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