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

Arne D. Hofemeier, Henning Hachmeister, Christian Pilger, Matthias Schürmann, Johannes F W Greiner, Lena Nolte, Holger Sudhoff, Christian Kaltschmidt, Thomas R Huser, Barbara Kaltschmidt

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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 PO4 3- 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.

Original languageEnglish (US)
Article number26716
JournalScientific Reports
Volume6
DOIs
StatePublished - May 26 2016
Externally publishedYes

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Raman Spectrum Analysis
Differentiation Antigens
Microscopy
Stem Cells
Durapatite
Turbinates
Adult Stem Cells
Bone Regeneration
Neural Crest
Tissue Engineering
Vibration
Osteoblasts
Osteogenesis
Cell Differentiation
Observation
Staining and Labeling
Bone and Bones
Polymerase Chain Reaction

ASJC Scopus subject areas

  • General

Cite this

Hofemeier, A. D., Hachmeister, H., Pilger, C., Schürmann, M., Greiner, J. F. W., Nolte, L., ... Kaltschmidt, B. (2016). Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. Scientific Reports, 6, [26716]. https://doi.org/10.1038/srep26716

Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. / Hofemeier, Arne D.; Hachmeister, Henning; Pilger, Christian; Schürmann, Matthias; Greiner, Johannes F W; Nolte, Lena; Sudhoff, Holger; Kaltschmidt, Christian; Huser, Thomas R; Kaltschmidt, Barbara.

In: Scientific Reports, Vol. 6, 26716, 26.05.2016.

Research output: Contribution to journalArticle

Hofemeier, AD, Hachmeister, H, Pilger, C, Schürmann, M, Greiner, JFW, Nolte, L, Sudhoff, H, Kaltschmidt, C, Huser, TR & Kaltschmidt, B 2016, 'Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells', Scientific Reports, vol. 6, 26716. https://doi.org/10.1038/srep26716
Hofemeier, Arne D. ; Hachmeister, Henning ; Pilger, Christian ; Schürmann, Matthias ; Greiner, Johannes F W ; Nolte, Lena ; Sudhoff, Holger ; Kaltschmidt, Christian ; Huser, Thomas R ; Kaltschmidt, Barbara. / Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells. In: Scientific Reports. 2016 ; Vol. 6.
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