Stem Cells
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First published online January 26, 2006
Stem Cells Vol. 24 No. 5 May 2006, pp. 1213 -1217
doi:10.1634/stemcells.2004-0324; www.StemCells.com
© 2006 AlphaMed Press

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TECHNOLOGY DEVELOPMENT

Metabolic Changes in Mesenchymal Stem Cells in Osteogenic Medium Measured by Autofluorescence Spectroscopy

Johann M.G. Reyesa, Sara Fermanianb, Fan Yangb, Shi-You Zhoua, Samantha Herretesa, Douglas B. Murphyc, Jennifer H. Elisseeffb, Roy S. Chucka

a Departments of Ophthalmology,
b Biomedical Engineering, and
c Cell Biology, Johns Hopkins University, Baltimore, Maryland, USA

Key Words. Bone marrow • Mesenchymal stem cells • Fluorescence microscopy • Differentiation

Correspondence: Roy S. Chuck, M.D., Ph.D., Wilmer Ophthalmological Institute, Johns Hopkins University, 3-127 Jefferson Building, 600 North Wolfe St., Baltimore, Maryland 21117, USA. Telephone: 410-502-1923; Fax: 443-287-1514; e-mail: rchuck1{at}jhmi.edu

Received on November 22, 2004; accepted for publication on December 14, 2005.

The purpose of this study was to measure metabolic changes in mesenchymal stem cells (MSCs) placed in osteogenic medium by autofluorescence spectroscopy. MSCs were plated in stem cell-supporting or osteogenic medium and imaged. Shift from the basic growth environment to the inductive osteogenic environment was confirmed by reverse transcription-polymerase chain reaction. Reduced pyridine nucleotides were detected by exciting near 366 nm and measuring fluorescence at 450 nm, and oxidized flavoproteins were detected by exciting at 460 nm and measuring fluorescence at 540 nm. The ratio of these fluorescence measurements, reduction-oxidation (redox) fluorometry, is a noninvasive measure of the cellular metabolic state. The detected pyridine nucleotide to flavoprotein ratio decreased upon transitioning from the stem cell to the differentiated state, as well as with increasing cell density and cell-cell contact. MSC metabolism increased upon placement in differentiating medium and with increasing cell density and contact. Redox fluorometry is a feasible, noninvasive technique for distinguishing MSCs from further differentiated cells.




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[Abstract] [Full Text] [PDF]


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