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

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 struc...

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Main Authors:	Donders, Raf, Sanen, Kathleen, Paesen, Rik, Slenders, Eli, Gyselaers, Wilfried, Stinissen, Piet, Ameloot, Marcel, Hellings, Niels
Format:	Online
Language:	English
Published:	Hindawi Publishing Corporation 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5056264/

id	pubmed-5056264
recordtype	oai_dc
spelling	pubmed-50562642016-10-16 Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells Donders, Raf Sanen, Kathleen Paesen, Rik Slenders, Eli Gyselaers, Wilfried Stinissen, Piet Ameloot, Marcel Hellings, Niels Research Article 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. Hindawi Publishing Corporation 2016 2016-09-26 /pmc/articles/PMC5056264/ /pubmed/27746820 http://dx.doi.org/10.1155/2016/5457132 Text en Copyright © 2016 Raf Donders et al. https://creativecommons.org/licenses/by/4.0/ 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.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Donders, Raf Sanen, Kathleen Paesen, Rik Slenders, Eli Gyselaers, Wilfried Stinissen, Piet Ameloot, Marcel Hellings, Niels
spellingShingle	Donders, Raf Sanen, Kathleen Paesen, Rik Slenders, Eli Gyselaers, Wilfried Stinissen, Piet Ameloot, Marcel Hellings, Niels Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
author_facet	Donders, Raf Sanen, Kathleen Paesen, Rik Slenders, Eli Gyselaers, Wilfried Stinissen, Piet Ameloot, Marcel Hellings, Niels
author_sort	Donders, Raf
title	Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
title_short	Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
title_full	Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
title_fullStr	Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
title_full_unstemmed	Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells
title_sort	label-free imaging of umbilical cord tissue morphology and explant-derived cells
description	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.
publisher	Hindawi Publishing Corporation
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5056264/
_version_	1613677692146679808

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

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