Characterization of human corneal stem cells by synchrotron infrared micro-spectroscopy.
OBJECTIVE
The purpose of this study was to use high resolution synchrotron radiation-based Fourier Transform Infrared (FTIR) micro-spectroscopy coupled with multivariate analysis to investigate the characteristics of adult stem cell (SC) and transit amplifying (TA) cell populations of the human corneal epithelium.


METHODS
Spectra of individual SC and TA cells in situ from cryosections of human cornea were collected using a synchrotron micro-spectroscopy facility at Daresbury laboratory, UK. Multivariate analysis and Mann Whitney U tests were used to analyse the spectral data from the SC and TA cell populations.


RESULTS
There were marked differences between the median spectra of the two cell populations. This correlated with their level of differentiation and functional specialization. Multivariate (principal component) analysis revealed that the cell populations could be segregated into distinct clusters, with only slight overlap between the two cell types. Significant (p<0.05) spectral differences were found in the spectral regions associated with nucleic acid, protein and lipids.


CONCLUSIONS
Synchrotron FTIR micro-spectroscopy together with principal component analysis is able to discriminate between SC and TA cell populations. Our results also suggest a small sub-population of corneal epithelial cells in the SC niche have TA cell-like characteristics. Many of the spectral differences between the SC and TA cell populations relate to differences in nucleic acid conformation.
Bentley
Adam J
AJ
Biomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Lancaster, UK. a.bentley@lancaster.ac.uk <a.bentley@lancaster.ac.uk>
Nakamura
Takahiro
T
Hammiche
Azzedine
A
Pollock
Hubert M
HM
Martin
Francis L
FL
Kinoshita
Shigeru
S
Fullwood
Nigel J
NJ
D000328:Adult; D000368:Aged; D002454:Cell Differentiation; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism; D008297:Male; D008875:Middle Aged; D008968:Molecular Conformation; D015999:Multivariate Analysis; D009696:Nucleic Acids / Q000737:chemistry / Q000378:metabolism; D025341:Principal Component Analysis; D017550:Spectroscopy, Fourier Transform Infrared; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014018:Tissue Distribution
D006023:Glycoproteins; D009696:Nucleic Acids