Growth substrate induced functional changes elucidated by FTIR and Raman spectroscopy in in–vitro cultured human keratinocytes

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• 作者：Aidan D. Meade ; Fiona M. Lyng ; Peter Knief and Hugh J. Byrne
• 关键词：Raman/FTIR spectroscopy ; HaCaT keratinocytes ; Adhesion effects ; Spectroscopic substrates
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2007
• 出版时间：March, 2007
• 年：2007
• 卷：387
• 期：5
• 页码：1717-1728
• 全文大小：559 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

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.