Document Detail


Growth substrate induced functional changes elucidated by FTIR and Raman spectroscopy in in-vitro cultured human keratinocytes.
MedLine Citation:
PMID:  17102969     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Non-invasive measurements of cellular function in in vitro cultured cell lines using vibrational spectroscopy require the use of spectroscopic substrates such as quartz, ZnSe and MirrIR etc. These substrates are generally dissimilar to the original in vivo extracellular environment of a given cell line and are often tolerated poorly by cultured cell lines resulting in morphological and functional changes in the cell. The present study demonstrates various correlations between vibrational spectroscopic analyses and biochemical analyses in the evaluation of the interaction of a normal human epithelial keratinocyte cell line (HaCaT) with MirrIR and quartz substrates coated with fibronectin, laminin and gelatin. The findings of this study suggest that there is a correlation between quantitative measurements of cellular proliferative capacity and viability and peak area ratios in FTIR spectra, with replicated differences in similar areas of the observed Raman spectra. Differences in the physiology of cells were observed between the two spectroscopic substrates coated in fibronectin and laminin, but little differences were observed when the cells were attached to gelatin-coated quartz and MirrIR slides. The correlations demonstrate the sensitivity of the spectroscopic techniques to evaluate the physiology of the system. Furthermore the study suggests that gelatin is a suitable coating for use in spectroscopic measurements of cellular function in human keratinocytes, as it provides a material that normalises the effect of substrate attachment on cellular physiology. This effect is likely to be cell-line dependent, and it is recommended that similar evaluations of this effect are performed for those combinations of spectroscopic substrate and cell lines that are to be used in individual experiments.
Authors:
Aidan D Meade; Fiona M Lyng; Peter Knief; Hugh J Byrne
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-11-11
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  387     ISSN:  1618-2642     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2007 Mar 
Date Detail:
Created Date:  2007-02-20     Completed Date:  2007-05-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1717-28     Citation Subset:  IM    
Affiliation:
School of Physics, Faculty of Science, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland. aidan.meade@dit.ie
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Cell Proliferation / drug effects
Humans
Intercellular Signaling Peptides and Proteins / administration & dosage*
Keratinocytes / drug effects*,  physiology*
Spectroscopy, Fourier Transform Infrared / methods*
Spectrum Analysis, Raman / methods*
Chemical
Reg. No./Substance:
0/Intercellular Signaling Peptides and Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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