Label-free discrimination analysis of de-differentiated vascular smooth
muscle cells, mesenchymal stem cells and their vascular and osteogenic
progeny using vibrational spectroscopy
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
Metadata
Item Type:
Article (Published)
Refereed:
Yes
Additional Information:
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.bbamcr.2017.11.006.
Uncontrolled Keywords:
Fourier transform infrared spectroscopy; Raman spectroscopy;
Stem cells; Myogenic differentiation; Linear discriminant analysis;
Principal component analysis
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), e Irish Research Council postgraduate scholarships to CM (GOIPG/2013/1144) and MDL (GOIPG/2014/43)
ID Code:
27811
Deposited On:
29 Sep 2022 13:13 by
Thomas Murtagh
. Last Modified 29 Sep 2022 13:13
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