COMPARISON OF 2D AND 3D PRIMARY CELL CULTURES OBTAINED FROM EXPLANT OF HIGH-GRADE UROTHELIAL BLADDER CANCER
DOI:
https://doi.org/10.15407/exp-oncology.2023.01.130Keywords:
apoptosis, bladder cancer, lactate dehydrogenase, primary cell culture, spheroidsAbstract
Studying the biological characteristics of bladder cancer in primary culture can be an effective way for diagnostic and prognostic purposes, as well as choosing a scheme for personalized therapy. Aim: To characterize and compare 2D and 3D primary cell cultures obtained from the same tumor sample resected from a patient with high-grade bladder cancer. Materials and Methods: 2D and 3D primary cell cultures were obtained from explants of resected bladder cancer. Glucose metabolism, lactate dehydrogenase (LDH) activity, and level of apoptosis were studied. Results: Multicellular tumor spheroids (3D) differ from planar culture (2D) by more pronounced consumption of glucose from the culture medium (1.7 times higher than 2D on Day 3 of culture), increased lactate dehydrogenase activity (2.5 times higher on Day 3 vs. Day 1 of cultivation, while in 2D culture LDH activity is constant), stronger acidification of the extracellular environment (pH dropped by 1 in 3D and by 0.5 in 2D). Spheroids demonstrate enhanced resistance to apoptosis (1.4 times higher). Conclusion: This methodological technique can be used both for tumor characterization and for selection of optimal postoperative chemotherapeutic schemes.
References
Habanjar O, Diab-Assaf M, Caldefie-Chezet F, Delort L. 3D cell culture systems: tumor application, advantages, and disadvantages. Int J Mol Sci 2021;22: 12200. doi: 10.3390/ijms222212200
Yoshida T, Okuyama H, Endo H, Inoue M. Spheroid cultures of primary urothelial cancer cells: cancer tissue-originated spheroid (CTOS) method. Methods Mol Biol 2018; 1655:145–153. doi: 10.1007/978-1-4939-7234-0_12
Driehuis E, Kretzschmar K, Clevers H. Establishment of patient-derived cancer organoids for drug-screening applications. Nat Protoc 2020; 15: 3380–3409. doi: 10.1038/s41596-020-0379-4
Nikolaienko TV, Nikulina VV, Shelest DV, et al. The mechanism of VEGF-mediated endothelial cells survival and proliferation in conditions of unfed-culture. Ukr Biochem J 2016; 88: 12–19. doi: 10.15407/ubj88.04.012
Freyer D, Harms C. Kinetic lactate dehydrogenase assay for detection of cell damage in primary neuronal cell cultures. Bio Protoc 2017; 7: e2308. doi: 10.21769/BioProtoc.2308
Wlodkowic D, Skommer J, Darzynkiewicz Z. Flow cytometry-based apoptosis detection. Methods Mol Biol 2009; 559: 19–32. doi: 10.1007/978-1-60327-017-5_2
Amin M. Histological variants of urothelial carcinoma: diagnostic, therapeutic and prognostic implications. Mod Pathol 2009; 22:S96–S118. doi: 10.1038/modpathol.2009.26
Alderson M, Grivas P, Milowsky MI, Wobker SE. Histologic variants of urothelial carcinoma: morphology, molecular features and clinical implications. Bladder Cancer 2020; 6: 107–122. doi: 10.3233/BLC-190257
Hirschhaeuser F, Sattler UG, Mueller-Klieser W. Lactate: a metabolic key player in cancer. Cancer Res 2011; 71: 6921–6925. doi: 10.1158/0008-5472.CAN-11-1457
Beckert S, Farrahi F, Aslam RS, et al. Lactate stimulates endothelial cell migration. Wound Repair Regen 2006; 14: 321-324.
Corbet C, Feron O. Tumour acidosis: From the passenger to the driver’s seat. Nat Rev Cancer2017; 17: 577–593. doi: 10.1038/nrc.2017.77
White KA, Grillo-Hill BK, Barber DL. Cancer cell behaviors mediated by dysregulated pH dynamics at a glance. J Cell Sci 2017; 130: 663–669. doi:10.1242/jcs.195297
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Experimental Oncology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.