Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells

Hofemeier AD, Hachmeister H, Pilger C, Schürmann M, Greiner J, Nolte L, Sudhoff H, Kaltschmidt C, Huser T, Kaltschmidt B (2016)
Scientific Reports 6: 26716.

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Abstract / Bemerkung
Tissue engineering by stem cell differentiation is a novel treatment option for bone regeneration. Most approaches for the detection of osteogenic differentiation are invasive or destructive and not compatible with live cell analysis. Here, non-destructive and label-free approaches of Raman spectroscopy, coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy were used to detect and image osteogenic differentiation of human neural crest-derived inferior turbinate stem cells (ITSCs). Combined CARS and SHG microscopy was able to detect markers of osteogenesis within 14 days after osteogenic induction. This process increased during continued differentiation. Furthermore, Raman spectroscopy showed significant increases of the PO43− symmetric stretch vibrations at 959 cm−1 assigned to calcium hydroxyapatite between days 14 and 21. Additionally, CARS microscopy was able to image calcium hydroxyapatite deposits within 14 days following osteogenic induction, which was confirmed by Alizarin Red-Staining and RT- PCR. Taken together, the multimodal label-free analysis methods Raman spectroscopy, CARS and SHG microscopy can monitor osteogenic differentiation of adult human stem cells into osteoblasts with high sensitivity and spatial resolution in three dimensions. Our findings suggest a great potential of these optical detection methods for clinical applications including in vivo observation of bone tissue–implant-interfaces or disease diagnosis.
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Scientific Reports
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6
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26716
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Article Processing Charge funded by the Deutsche Forschungsgemeinschaft and the Open Access Publication Fund of Bielefeld University.
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Hofemeier AD, Hachmeister H, Pilger C, et al. Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports. 2016;6: 26716.
Hofemeier, A. D., Hachmeister, H., Pilger, C., Schürmann, M., Greiner, J., Nolte, L., Sudhoff, H., et al. (2016). Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports, 6, 26716. doi:10.1038/srep26716
Hofemeier, A. D., Hachmeister, H., Pilger, C., Schürmann, M., Greiner, J., Nolte, L., Sudhoff, H., Kaltschmidt, C., Huser, T., and Kaltschmidt, B. (2016). Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports 6:26716.
Hofemeier, A.D., et al., 2016. Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports, 6: 26716.
A.D. Hofemeier, et al., “Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells”, Scientific Reports, vol. 6, 2016, : 26716.
Hofemeier, A.D., Hachmeister, H., Pilger, C., Schürmann, M., Greiner, J., Nolte, L., Sudhoff, H., Kaltschmidt, C., Huser, T., Kaltschmidt, B.: Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports. 6, : 26716 (2016).
Hofemeier, Arne D., Hachmeister, Henning, Pilger, Christian, Schürmann, Matthias, Greiner, Johannes, Nolte, Lena, Sudhoff, Holger, Kaltschmidt, Christian, Huser, Thomas, and Kaltschmidt, Barbara. “Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells”. Scientific Reports 6 (2016): 26716.
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4 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Fast epi-detected broadband multiplex CARS and SHG imaging of mouse skull cells.
Capitaine E, Moussa NO, Louot C, Bardet SM, Kano H, Duponchel L, Lévêque P, Couderc V, Leproux P., Biomed Opt Express 9(1), 2018
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Masia F, Glen A, Stephens P, Langbein W, Borri P., J Biophotonics 11(7), 2018
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