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Three-dimensional cell spheroid culture and cell viability study of uveal melanoma cell line C918 with luteolin treatment.

Authors :
Jinhai Y
Yunxiu C
Qi J
Jiancheng G
Zhida P
Sha W
Hongfei L
Qihua X
Source :
International ophthalmology [Int Ophthalmol] 2024 Sep 22; Vol. 44 (1), pp. 385. Date of Electronic Publication: 2024 Sep 22.
Publication Year :
2024

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.)

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