Evaluation of Melanoma (SK-MEL-2) Cell Growth between Three-Dimensional (3D) and Two-Dimensional (2D) Cell Cultures with Fourier Transform Infrared (FTIR) Microspectroscopy

• Published in: International journal of molecular sciences Vol. 21; no. 11; p. 4141
• Main Authors: Srisongkram, Tarapong, Weerapreeyakul, Natthida, Thumanu, Kanjana
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 10.06.2020; MDPI
• Subjects: Apoptosis; Cell culture; Cell Culture Techniques - methods; Cell Death; Cell Line, Tumor; Cell Proliferation; Cluster analysis; colony formed cells; Deoxyribonucleic acid; DNA; Fluorescence; Fourier analysis; Fourier transform infrared microspectroscopy (FTIR); Humans; Infrared spectroscopy; Integrals; Lipids; Lipids - chemistry; Melanoma; Melanoma - chemistry; Melanoma - genetics; Melanoma - pathology; Multivariate analysis; Principal Component Analysis; Principal components analysis; Protein structure; Proteins; Proteins - chemistry; RNA, Neoplasm - chemistry; Secondary structure; Skin cancer; Spectroscopy, Fourier Transform Infrared; Spectrum analysis; spheroid cell culture; Spheroids; Spheroids, Cellular - pathology; Staining; three-dimensional (3D) cell culture; spheroid cell culture; melanoma; Fourier transform infrared microspectroscopy (FTIR); colony formed cells; multivariate analysis; three-dimensional (3D) cell culture
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21114141

Fourier transform infrared (FTIR) microspectroscopy was used to evaluate the growth of human melanoma cells (SK-MEL-2) in two-dimensional (2D) versus three-dimensional (3D) spheroid culture systems. FTIR microspectroscopy, coupled with multivariate analysis, could be used to monitor the variability of spheroid morphologies prepared from different cell densities. The characteristic shift in absorbance bands of the 2D cells were different from the spectra of cells from 3D spheroids. FTIR microspectroscopy can also be used to monitor cell death similar to fluorescence cell staining in 3D spheroids. A change in the secondary structure of protein was observed in cells from the 3D spheroid versus the 2D culture system. FTIR microspectroscopy can detect specific alterations in the biological components inside the spheroid, which cannot be detected using fluorescence cell death staining. In the cells from 3D spheroids, the respective lipid, DNA, and RNA region content represent specific markers directly proportional to the spheroid size and central area of necrotic cell death, which can be confirmed using unsupervised PCA and hierarchical cluster analysis. FTIR microspectroscopy could be used as an alternative tool for spheroid cell culture discrimination, and validation of the usual biochemical technique.