Molony, Claire and McIntyre, Jennifer and Maguire, Adrian and Hakimjavadi, Roya and Burtenshaw, Denise and Casey, Gillian and Luca, Mariana Di and Hennelly, Bryan M. and Byrne, Hugh J. and Cahill, Paul A. (2018) Label-free discrimination analysis of de-differentiated vascular smooth muscle cells, mesenchymal stem cells and their vascular and osteogenic progeny using vibrational spectroscopy. BBA - Molecular Cell Research, 1865. pp. 343-353. ISSN 0167-4889

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Abstract

The accumulation of vascular smooth muscle (SMC)-like cells and stem cell-derived myogenic and osteogenic progeny contributes significantly to arteriosclerotic disease. This study established whether label-free vibrational spectroscopy can discriminate de-differentiated ‘synthetic’ SMCs from undifferentiated stem cells and their myogenic and osteogenic progeny in vitro, compared with conventional immunocytochemical and genetic analyses. TGF-β1- and Jagged1-induced myogenic differentiation of CD44+ mesenchymal stem cells was confirmed in vitro by immunocytochemical analysis of specific SMC differentiation marker expression (α-actin, calponin and myosin heavy chain 11), an epigenetic histone mark (H3K4me2) at the myosin heavy chain 11 locus, promoter transactivation and mRNA transcript levels. Osteogenic differentiation was confirmed by alizarin red staining of calcium deposition. Fourier Transform Infrared (FTIR) maps facilitated initial screening and discrimination while Raman spectroscopy of individual cell nuclei revealed specific spectral signatures of each cell type in vitro, using Principal Components Analysis (PCA). PCA fed Linear Discriminant Analysis (LDA) enabled quantification of this discrimination and the sensitivity and specificity value was determined for all cell populations based on a leave-one-out cross validation method and revealed that de-differentiated SMCs and stem-cell derived myogenic progeny in culture shared the greatest similarity. FTIR and Raman spectroscopy discriminated undifferentiated stem cells from both their myogenic and osteogenic progeny. The ability to detect stem cell-derived myogenic progeny using label-free platforms in situ may facilitate interrogation of these important phenotypes during vascular disease progression.

Item Type:	Article
Additional Information:	This work was supported by research grants from Science Foundation Ireland to PAC (SFI 11/PI/1128), HJB (SFI PI 11/PI/08) and BH (SFI 11/SIRG/I2140 and 15/CDA/366) and the Irish Research Council postgraduate scholarships to CM (GOIPG/2013/1144) and MDL (GOIPG/2014/43).
Keywords:	Fourier transform infrared spectroscopy; Raman spectroscopy; Stem cells; Myogenic differentiation; Linear discriminant analysis; Principal component analysis;
Academic Unit:	Faculty of Science and Engineering > Computer Science Faculty of Science and Engineering > Electronic Engineering
Item ID:	13132
Identification Number:	https://doi.org/10.1016/j.bbamcr.2017.11.006
Depositing User:	Dr. Bryan Hennelly
Date Deposited:	22 Jul 2020 14:17
Journal or Publication Title:	BBA - Molecular Cell Research
Publisher:	Elsevier
Refereed:	Yes
Funders:	Science Foundation Ireland (SFI), Irish Research Council (IRC)
URI:	Publisher
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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