title: Proton NMR characterization of intact primary and metastatic melanoma cells in 2D & 3D cultures.
abstract: OBJECTIVE
  To characterize the differences between the primary and metastatic melanoma cell lines grown in 2D cultures and 3D cultures.
  
  
  METHODS
  Primary melanoma cells (WM115) and metastatic melanoma cells (WM266) extracted from a single donor was cultured in 2D as well as 3D cultures. These cells were characterized using proton NMR spectrometry, and the qualitative chemical shifts markers were identified and discussed.
  
  
  RESULTS
  In monolayer culture (2D), we observed one qualitative chemical shift marker for primary melanoma cells. In spheroid cultures (3D), we observed nine significant chemical shifts, of which eight markers were specific for primary melanoma spheroids, whereas the other one marker was specific to metastatic melanoma spheroids. This study suggests that the glucose accumulation and phospholipid composition vary significantly between the primary and metastatic cells lines that are obtained from a single donor and also with the cell culturing methods. 14 qualitative chemical shift markers were obtained in the comparison between monolayer culture and spheroids cultures irrespective of the differences in the cell lines. Among which 4 were unique to monolayer cultures whereas 10 chemical shifts were unique to the spheroid cultures. This study also shows that the method of cell culture would drastically affect the phospholipid composition of the cells and also depicts that the cells in spheroid culture closely resembles the cells in vivo.
  
  
  CONCLUSIONS
  This study shows the high specificity of proton NMR spectrometry in characterizing cancer cell lines and also shows the variations in the glucose accumulation and phospholipid composition between the primary and metastatic melanoma cell lines from the same donor. Differences in the cell culture method does plays an important role in phospholipid composition of the cells.
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, E1-08-016, 9 Engineering Drive 1, Singapore, 117575, Singapore.
authors.forename: Gokula Krishnan
authors.identifier: http://orcid.org/0000-0001-5365-4348
authors.initials: GK
authors.lastname: Ramachandran
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, E1-08-016, 9 Engineering Drive 1, Singapore, 117575, Singapore. bieych@nus.edu.sg.
authors.forename: Chen Hua
authors.initials: CH
authors.lastname: Yeow
mesh_terms: D000704:Analysis of Variance; D014408:Biomarkers, Tumor; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008545:Melanoma / Q000378:metabolism / Q000473:pathology* / Q000556:secondary*; D010743:Phospholipids / Q000032:analysis / Q000378:metabolism; D018874:Spheroids, Cellular; D013997:Time Factors
chemical_list: D014408:Biomarkers, Tumor; D010743:Phospholipids; D005947:Glucose
keywords: Cancer; Melanoma; Nuclear magnetic resonance; Proton magnetic resonance spectrometry