Development of Spheroid in Co-Culture of Prostate Tumor with Human Fibroblast, using the Hanging Drop Technique with Plate Inversion for Analysis of GammaCell220 Irradiation in Co-60
DOI:
https://doi.org/10.15392/2319-0612.2024.2682Keywords:
Hanging Drop, LNCaP, GammaCell Co-60Abstract
The development of tumor spheroids in co-culture with human fibroblasts represents an innovative approach to modeling the prostate cancer microenvironment and assessing radiation effects. In this study, we employed the "hanging drop" technique in conjunction with plate inversion to generate three-dimensional spheroids composed of prostate tumor cells (LNCAP cell line) co-cultivated with human fibroblasts. This approach aims to create a more representative in vivo-like tumor environment, facilitating a detailed analysis of the interactions between tumor cells and fibroblasts in 3D conditions. The formed spheroids were exposed to gamma radiation using a GammaCell 220 irradiator with Co-60, with the goal of investigating the effects of radiation on both fibroblastic and tumor cells. Gamma radiation is known to induce cell death, and this study examines how such damage impacts cellular morphology within the spheroid microenvironment.
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[1] BOLTE, S.; CORDELIÈRES, F. P. A guided tour into subcellular colocalization analysis in light microscopy. Journal of Microscopy, v. 224, p. 213–232, 2007. DOI: https://doi.org/10.1111/j.1365-2818.2006.01706.x
[2] CHAICHAROENAUDOMRUNG, N.; KUNHORM, P.; NOISA, P. Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling. World Journal of Stem Cells, v. 11, n. 12, p. 1065–1076, 2019. DOI: https://doi.org/10.4252/wjsc.v11.i12.1065
[3] EILENBERGER, C. et al. Effect of spheroidal age on sorafenib diffusivity and toxicity in a 3D HepG2 spheroid model. Scientific Reports, v. 9, n. 1, p. 1–10, 2019. DOI: https://doi.org/10.1038/s41598-019-41273-3
[4] HORMAN, S. R. et al. 3D high-content analysis of spheroids. Genetic Engineering & Biotechnology News, v. 33, n. 16, p. 18–19, 2013. DOI: https://doi.org/10.1089/gen.33.16.08
[5] IVANOV, D. P. et al. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres. PLOS ONE, v. 9, n. 8, p. e103817, 2014. DOI: https://doi.org/10.1371/journal.pone.0103817
[6] IVOSEV, G.; BURTON, L.; BONNER, R. F. Dimensionality reduction and visualization in principal component analysis. Analytical Chemistry, v. 80, n. 13, p. 4933–4944, 2008. DOI: https://doi.org/10.1021/ac800110w
[7] JENSEN, C.; TENG, Y. Is it time to start transitioning from 2D to 3D cell culture? Frontiers in Molecular Biosciences, v. 7, p. 33, 2020. DOI: https://doi.org/10.3389/fmolb.2020.00033
[8] JOHNSON, R. J.; CARRINGTON, B. M. Pelvic radiation disease. Clinical Radiology, v. 45, n. 1, p. 4–10, 1992. DOI: https://doi.org/10.1016/S0009-9260(05)81458-8
[9] MEHTA, G. et al. Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. Journal of Controlled Release, v. 164, n. 2, p. 192–204, 2012. DOI: https://doi.org/10.1016/j.jconrel.2012.04.045
[10] MITTLER, F. et al. High-content monitoring of drug effects in a 3D spheroid model. Frontiers in Oncology, v. 7, p. 293, 2017. DOI: https://doi.org/10.3389/fonc.2017.00293
[11] MURÓYA, Y. et al. High-LET ion radiolysis of water: visualization of the formation and evolution of ion tracks and relevance to the radiation-induced bystander effect. Radiation Research, v. 165, n. 4, p. 485–491, 2006. DOI: https://doi.org/10.1667/RR3540.1
[12] PANGANIBAN, R. A. M.; SNOW, A. L.; DAY, R. M. Mechanisms of radiation toxicity in transformed and non-transformed cells. International Journal of Molecular Sciences, v. 14, n. 8, p. 15931–15959, 2013. DOI: https://doi.org/10.3390/ijms140815931
[13] RUA, Y. et al. Correlation between cellular uptake and cytotoxicity of polystyrene micro/nanoplastics in HeLa cells: a size-dependent matter. PLOS ONE, v. 18, n. 8, p. e0289473, 2023. DOI: https://doi.org/10.1371/journal.pone.0289473
[14] SART, S. et al. Multiscale cytometry and regulation of 3D cell cultures on a chip. Nature Communications, v. 8, p. 475, 2017. DOI: https://doi.org/10.1038/s41467-017-00475-x
[15] SGOUROS, G.; YANG, W.-H.; ENMON, R. M. Spheroids of prostate tumor cell lines. In: Spheroids of Prostate Tumor Cell Lines. New Jersey: Humana Press, 2003. p. 79–90. DOI: https://doi.org/10.1385/1-59259-372-0:79
[16] ZANONI, M. et al. Anticancer drug discovery using multicellular tumor spheroid models. Expert Opinion on Drug Discovery, v. 14, n. 3, p. 289–303, 2019. DOI: https://doi.org/10.1080/17460441.2019.1570129
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Copyright (c) 2026 Thais Monteiro, Alex Alves Rodrigues, Giovana Dias da Silva, Esther Carolina dos Santos, Patricia Lima Falcão, Suleyna Ribeiro Prudente, Mayelle Maria Paz, Daniel Perez Vieira
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