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Three-dimensional cell spheroid culture and cell viability study of uveal melanoma cell line C918 with luteolin treatment.
- Source :
-
International ophthalmology [Int Ophthalmol] 2024 Sep 22; Vol. 44 (1), pp. 385. Date of Electronic Publication: 2024 Sep 22. - Publication Year :
- 2024
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Abstract
- Objective: This study aims to investigate the morphological and histological characteristics of three-dimensional cell spheroids derived from the uveal melanoma (UM) cell line C918 and assess the impact of luteolin on their cell viability.<br />Methods: C918 cells were cultured in ultra-low adsorption 96-well plates, and morphological changes in C918 three-dimensional cell spheroids were observed over varying time intervals. Histological features of C918 multicellular spheroids cultured in ultra-low adsorption 6-well plates were examined using both HE staining and immunohistochemical staining. The CCK8 reagent was employed to measure the optical density at a 450 nm wavelength after 72-h treatments with varying luteolin concentrations in both two-dimensional and three-dimensional cultured C918 cells. The IC50 values were compared between the two culture conditions.<br />Results: Over time in culture, the volume of C918 three-dimensional cell spheroids gradually increased, and an ischemic- and hypoxic-like region became evident within the spheroids on days 4 to 6 of culture. Histological staining demonstrated positive expression of cell viability marker antibodies (Ki67) and melanoma marker antibodies (MelanA, HMB45, S-100) in the multicellular spheroids from three-dimensional culture. CCK-8 experiments revealed that the IC50 values for luteolin in C918 cells were 183.50 μmol/L in three-dimensional culture and 16.19 μmol/L in two-dimensional culture after 72 h. Three-dimensional cultured C918 cells, treated with varying luteolin concentrations for 72 h, were observed under a microscope. The maximum cross-sectional area showed no statistically significant differences between the groups, but it was reduced in comparison to the control group.<br />Conclusion: Three-dimensional cultured C918 cell spheroids exhibit histological characteristics similar to real tumors and are less responsive to luteolin than their two-dimensional counterparts. They offer a valuable model for anti-tumor drug screening.<br /> (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)
- Subjects :
- Humans
Tumor Cells, Cultured
Cell Line, Tumor
Cell Proliferation drug effects
Cell Culture Techniques, Three Dimensional methods
Luteolin pharmacology
Uveal Neoplasms drug therapy
Uveal Neoplasms pathology
Spheroids, Cellular
Melanoma drug therapy
Melanoma pathology
Cell Survival drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1573-2630
- Volume :
- 44
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- International ophthalmology
- Publication Type :
- Academic Journal
- Accession number :
- 39306600
- Full Text :
- https://doi.org/10.1007/s10792-024-03312-5