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

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

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.

Original languageEnglish
Title of host publicationOptics in Health Care and Biomedical Optics VII
EditorsYuguo Tang, Xingde Li, Ying Gu, Dan Zhu, Qingming Luo
PublisherSPIE
ISBN (Electronic)9781510604674
DOIs
StatePublished - 2016
EventOptics in Health Care and Biomedical Optics VII - Beijing, China
Duration: 12 Oct 201614 Oct 2016

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume10024
ISSN (Print)0277-786X
ISSN (Electronic)1996-756X

Conference

ConferenceOptics in Health Care and Biomedical Optics VII
Country/TerritoryChina
CityBeijing
Period12/10/1614/10/16

Keywords

  • adipogenesis
  • cellular differentiation
  • Fluorescence lifetime imaging microscopy (FLIM)
  • human mesenchymal stem cells (hMSCs)
  • NADH autofluorescence
  • osteogenesis
  • reduced nicotinamide adenine dinucleotide (NADH)

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