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Stem Cells International
Volume 2016, Article ID 5457132, 15 pages
http://dx.doi.org/10.1155/2016/5457132
Research Article

Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells

1Biomedical Research Institute, Hasselt University and School of Life Sciences, Transnational University Limburg, Agoralaan Building C, 3590 Diepenbeek, Belgium
2Ziekenhuis Oost-Limburg, Campus St. Jan, Schiepse Bos 6, 3600 Genk, Belgium

Received 14 May 2016; Revised 28 July 2016; Accepted 31 July 2016

Academic Editor: Francesco Petrella

Copyright © 2016 Raf Donders et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

In situ detection of MSCs remains difficult and warrants additional methods to aid with their characterization in vivo. Two-photon confocal laser scanning microscopy (TPM) and second harmonic generation (SHG) could fill this gap. Both techniques enable the detection of cells and extracellular structures, based on intrinsic properties of the specific tissue and intracellular molecules under optical irradiation. TPM imaging and SHG imaging have been used for label-free monitoring of stem cells differentiation, assessment of their behavior in biocompatible scaffolds, and even cell tracking in vivo. In this study, we show that TPM and SHG can accurately depict the umbilical cord architecture and visualize individual cells both in situ and during culture initiation, without the use of exogenously applied labels. In combination with nuclear DNA staining, we observed a variance in fluorescent intensity in the vessel walls. In addition, antibody staining showed differences in Oct4, αSMA, vimentin, and ALDH1A1 expression in situ, indicating functional differences among the umbilical cord cell populations. In future research, marker-free imaging can be of great added value to the current antigen-based staining methods for describing tissue structures and for the identification of progenitor cells in their tissue of origin.