Three-dimensional Cell Culture Model for In-Vitro Drug-Testing Platforms in Cancer Research

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Prin Sungwan
Wunchana Seubwai


Chemotherapy and radiotherapy are widely used as a standard treatment regimen for many cancers. However, resistance to the standard treatment regimens usually occurs for several malignant tumors. Thus, searching for novel anticancer agents with high therapeutic effectiveness and less side effects is urgently needed. However, there are several evidences suggesting that two-dimensional (2D) cell culture models cannot recapitulate key characteristics of in-vivo tumor and do not provide in-vivo and clinically relevant data for anti-cancer drug discovery and development. Therefore, three-dimensional (3D) cell culture models have been developed. Cultured cells under 3D cell culture models provide several aspects of in-vivo solid tumor and offer better in-vitro models for anti-cancer drug screening and validation. This review article will clarify the limitations of 2D cell culture models and application of 3D cell culture models as a better in-vitro models for cancer research. Including, application 3D cell culture models in chemo- and radiotherapy drug testing, cancer metastasis, cancer stem cell and cancer metabolisms.


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Sungwan ป, Seubwai ว. Three-dimensional Cell Culture Model for In-Vitro Drug-Testing Platforms in Cancer Research. SRIMEDJ [Internet]. 2021 Oct. 4 [cited 2022 May 16];36(5):625-33. Available from:
Review Articles


1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018; 68(6): 394–424.
2. Avorn J. The $2.6 Billion Pill — Methodologic and Policy Considerations. N Engl J Med 2015; 372(20): 1877–1879.
3. Barnes PJ, Bonini S, Seeger W, Belvisi MG, Ward B, Holmes A. Barriers to new drug development in respiratory disease. Eur Respir J 2015; 45(5): 1197–1207.
4. Barbone D, Van Dam L, Follo C, Jithesh PV, Zhang SD, Richards WG, et al. Analysis of gene expression in 3D spheroids highlights a survival role for ASS1 in Mesothelioma. PLoS One 2016 ; 11(3): e0150044.
5. Yang Y, Roy A, Zhao Y, Undzys E, Li SD. Comparison of tumor penetration of podophyllotoxin–carboxymethylcellulose conjugates with various chemical compositions in tumor spheroid sulture and In vivo solid tumor. Bioconjug Chem 2017; 28(5): 1505–1518.
6. Langhans SA. Three-dimensional in vitro cell culture models in drug discovery and drug repositioning. Front Pharmacol [Internet]. 2018 [cited 2019 Oct 2];9. Available from:
7. In Vivo vs. In Vitro: definition, examples, and More [Internet]. [cited Dec 14, 2019]. Available from:
8. Allen DD, Caviedes R, Cárdenas AM, Shimahara T, Segura-Aguilar J, Caviedes PA. Cell lines as In vitro models for drug screening and toxicity studies. Drug Dev Ind Pharm 2005 ; 31(8): 757–768.
9. Katt ME, Placone AL, Wong AD, Xu ZS, Searson PC. In vitro tumor models: advantages, disadvantages, variables, and selecting the right platform. Front Bioeng Biotechnol [Internet]. 2016 Feb 12 [cited May 7, 2019];4. Available from:
10. Meijer TG, Naipal KA, Jager A, van Gent DC. Ex vivo tumor culture systems for functional drug testing and therapy response prediction. Future Sci OA 2017; 3(2): FSO190.
11. Introduction. Harvey, William. 1909-1914. On the motion of the heart and blood in animals. The Harvard Classics [Internet]. [cited May 4, 2019]; Available from:
12. Merten O-W. Advances in cell culture: anchorage dependence. Philos Trans R Soc Lond B Biol Sci [Internet]. 2015 Feb 5 [cited May 7, 2019];370(1661). Available from:
13. Nath S, Devi GR. Three-dimensional culture systems in cancer research: Focus on tumor spheroid model. Pharmacol Ther 2016; 163: 94–108.
14. Vinci M, Gowan S, Boxall F, Patterson L, Zimmermann M, Court W, et al. Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation. BMC Biol 2012;10: 29. doi: 10.1186/1741-7007-10-29.
15. Weiswald L-B, Bellet D, Dangles-Marie V. Spherical cancer models in tumor biology. Neoplasia 2015; 17(1): 1–15.
16. Edmondson R, Broglie JJ, Adcock AF, Yang L. Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. ASSAY Drug Dev Technol 2014 ; 12(4): 207–218.
17. Birgersdotter A, Sandberg R, Ernberg I. Gene expression perturbation in vitro—A growing case for three-dimensional (3D) culture systems. Semin Cancer Biol 2005; 15(5): 405–412.
18. Hutchinson L, Kirk R. High drug attrition rates—where are we going wrong?. Nat Rev Clin Oncol 2011; 8(4): 189–190.
19. Sutherland RM, McCredie JA, Inch WR. Growth of multicell spheroids in tissue culture as a model of nodular carcinomas. J Natl Cancer Inst 1971; 46(1): 113–120.
20. Holliday DL, Brouilette KT, Markert A, Gordon LA, Jones JL. Novel multicellular organotypic models of normal and malignant breast: tools for dissecting the role of the microenvironment in breast cancer progression. Breast Cancer Res 2009; 11(1): R3.
21. Sandberg R, Ernberg I. The molecular portrait of in vitro growth by meta-analysis of gene-expression profiles. Genome Biol 2005; 6(8): R65.
22. Delarue M, Montel F, Vignjevic D, Prost J, Joanny J-F, Cappello G. Compressive stress inhibits proliferation in tumor spheroids through a volume limitation. Biophys J 2014; 107(8): 1821–1828.
23. Cukierman E, Pankov R, Stevens DR, Yamada KM. Taking cell-matrix adhesions to the third dimension. Science 2001; 294(5547): 1708–1712.
24. Jung H-R, Kang HM, Ryu JW, Kim DS, Noh KH, Kim ES, et al. Cell spheroids with enhanced aggressiveness to mimic human liver cancer In vitro and In vivo. Sci Rep 2017; 7(1): 10499.
25. Costa EC, Moreira AF, de Melo-Diogo D, Gaspar VM, Carvalho MP, Correia IJ. 3D tumor spheroids: an overview on the tools and techniques used for their analysis. Biotechnol Adv 2016; 34(8): 1427–1441.
26. Tibbitt MW, Anseth KS. Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechnol Bioeng 2009; 103(4): 655–663.
27. Lovitt CJ, Shelper TB, Avery VM. Doxorubicin resistance in breast cancer cells is mediated by extracellular matrix proteins. BMC Cancer 2018; 18: 41.
28. Pickl M, Ries CH. Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab. Oncogene 2009; 28(3): 461–468.
29. Dubessy C, Merlin J-L, Marchal C, Guillemin F. Spheroids in radiobiology and photodynamic therapy. Critical reviews in Oncology/Hematology 2000; 36(2): 179–192.
30. Schwachöfer JH. Multicellular tumor spheroids in radiotherapy research (review). Anticancer Res 1990; 10(4): 963–965.
31. Terasima T, Tolmach LJ. Changes in x-ray sensitivity of HeLa cells during the division cycle. Nature 1961; 190: 1210–1211.
32. Olive PL, Durand RE. Drug and radiation resistance in spheroids: cell contact and kinetics. Cancer Metastasis Rev 1994; 13(2): 121–138.
33. Berens EB, Holy JM, Riegel AT, Wellstein A. A cancer cell spheroid assay to assess invasion in a 3D setting. J Vis Exp 2015; (105): 53409.
34. Lee JW, Sung JS, Park YS, Chung S, Kim YH. Isolation of spheroid-forming single cells from gastric cancer cell lines: enrichment of cancer stem-like cells. BioTechniques 2018; 65(4): 197–203.
35. Yu Z, Pestell TG, Lisanti MP, Pestell RG. Cancer stem cells. Int J Biochem Cell Biol 2012 ; 44(12): 2144–2151.
36. Guo X, Chen Y, Ji W, Chen X, Li C, Ge R. Enrichment of cancer stem cells by agarose multi-well dishes and 3D spheroid culture. Cell Tissue Res 2019; 375(2): 397–408.
37. Lee JW, Sung JS, Park YS, Chung S, Kim YH. Isolation of spheroid-forming single cells from gastric cancer cell lines: enrichment of cancer stem-like cells. BioTechniques 2018; 65(4): 197–203.
38. Ishiguro T, Ohata H, Sato A, Yamawaki K, Enomoto T, Okamoto K. Tumor‐derived spheroids: Relevance to cancer stem cells and clinical applications. Cancer Sci 2017; 108(3): 283–289.
39. Warburg O. On the Origin of Cancer Cells. Science 1956; 123(3191): 309–314.
40. Schwartz L, Supuran CT, Alfarouk KO. The Warburg Effect and the hallmarks of cancer. Anticancer Agents Med Chem 2017; 17(2): 164–170.
41. Koshkin V, Ailles LE, Liu G, Krylov SN. Metabolic suppression of a drug-resistant subpopulation in cancer spheroid Cells. J Cell Biochem 2016; 117(1): 59–65.
42. Dwarakanath B, Singh D, Banerji A, Sarin R, Venkataramana N, Jalali R, et al. Clinical studies for improving radiotherapy with 2-deoxy-D-glucose: Present status and future prospects. J Can Res Ther 2009; 5(9): 21.
43. Gaebler M, Silvestri A, Haybaeck J, Reichardt P, Lowery CD, Stancato LF, et al. Three-dimensional patient-derived In vitro sarcoma models: promising tools for improving clinical tumor management. Front Oncol 2017; 7: 203.
44. Verjans ET, Doijen J, Luyten W, Landuyt B, Schoofs L. Three-dimensional cell culture models for anticancer drug screening: Worth the effort?. J Cellular Physiol 2018; 233(4): 2993–3003.