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Archives of Medical Science
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vol. 14
State of the art paper

2D and 3D cell cultures – a comparison of different types of cancer cell cultures

Marta Kapałczyńska, Tomasz Kolenda, Weronika Przybyła, Maria Zajączkowska, Anna Teresiak, Violetta Filas, Matthew Ibbs, Renata Bliźniak, Łukasz Łuczewski, Katarzyna Lamperska

Arch Med Sci 2018; 14, 4: 910–919
Online publish date: 2016/11/18
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Cell culture is a widely used in vitro tool for improving our understanding of cell biology, tissue morphology, and mechanisms of diseases, drug action, protein production and the development of tissue engineering. Most research regarding cancer biology is based on experiments using two-dimensional (2D) cell cultures in vitro. However, 2D cultures have many limitations, such as the disturbance of interactions between the cellular and extracellular environments, changes in cell morphology, polarity, and method of division. These disadvantages led to the creation of models which are more closely able to mimic conditions in vivo. One such method is three-dimensional culture (3D). Optimisation of the culture conditions may allow for a better understanding of cancer biology and facilitate the study of biomarkers and targeting therapies. In this review, we compare 2D and 3D cultures in vitro as well as different versions of 3D cultures.

co-culture, cell culture methods, 3D culture, 2D culture, cancer research

Yamada K, Cukierman E. Modeling tissue morphogenesis and cancer in 3D. Cell 2007; 130: 601-10.
Harrison R. Observations of the living developing nerve fiber. Anat Rec 1907; 1: 116-28.
Harrison RG. The outgrowth of the nerve fiber as a mode of protoplasmic movement. J Exp Zool 1910; 9: 787-846.
Scudiero D, Shoemaker RH, Paull KD, et al. Evaluation of a soluble tetrazolium formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 1988; 48: 4827-33.
Jacoby W, Pasten I. Methods in Enzymology: Cell Culture. Vol. 58. Academic Press, New York 1979.
Ryan J. Introduction to Animal Cell Culture. Technical Bulletin Corning 2003; 1-8.
Sanyal S. Culture and assay systems used for 3D cell culture. Corning 2014; 9: 1-18.
Aggarwal B, Danda D, Gupta S, Gehlot P. Models for prevention and treatment of cancer: problems vs promises. Biochem Pharmacol 2009; 78: 1083-94.
Breslin S, O’Driscoll L. Three-dimensional cell culture: the missing link in drug discovery. Drug Discov Today 2013; 18: 240-9.
Pampaloni F, Reynaud EG, Stelzer EH. The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol 2007; 8: 839-45.
Baker B, Chen C. Deconstructing the third dimension – how 3D culture microenvironments alter cellular cues. J Cell Sci 2012; 125: 3015-24.
Hickman JA, Graeser R, de Hoogt R, et al. Three-dimensional models of cancer for pharmacology and cancer cell biology: capturing tumor complexity in vitro/ex vivo. Biotechnol J 2014; 9: 1115-28.
Bissell MJ, Rizki A, Mian IS. Tissue architecture: the ultimate regulator of breast epithelial function. Curr Opin Cell Biol 2003; 15: 753-62.
von der Mark K, Gauss V, von der Mark H, Müller P. Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature 1977; 267: 531-2.
Petersen OW, Rønnov-Jessen L, Howlett AR, Bissel MJ. Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA 1992; 89: 9064-8.
Mahmud G, Campbell CJ, Bishop KJM, et al. Directing cell motions on micropatterned ratchets. Nat Phys 2009; 5: 606-12.
Kilian K, Bugarija B, Lahn BT, et al. Geometric cues for directing the differentiation of mesenchymal stem cells. Proc Natl Acad Sci USA 2010; 107: 4872-7.
Debnath J, Brugge JS. Modelling glandular epithelial cancers in three-dimensional cultures. Nat Rev Cancer 2005; 5: 675-88.
Nelson CM, Bissell MJ. Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Annu Rev Cell Dev Biol 2006; 22: 287-309.
Mseka T, Bamburg JR, Cramer LP. ADF/cofilin family proteins control formation of oriented actin-filament bundles in the cell body to trigger fibroblast polarization. J Cell Sci 2007; 120: 4332-44.
Weaver V, Lelièvre S, Lakins JN, et al. Beta4 integrin-dependent formation of polarized three-dimensional architecture confers resistance to apoptosis in normal and malignant mammary epithelium. Cancer Cell 2002; 2: 205-16.
Meyers J, Craig J, Odde DJ. Potential for control of signaling pathways via cell size and shape. Curr Biol 2003; 16: 1685-93.
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: 405-12.
Li C, Kato M, Shiue L, Shively JE, Ares M, Lin RJ. Cell type and culture condition-dependent alternative splicing in human breast cancer cells revealed by splicing-sensitive microarrays. Cancer Res 2006; 66: 1990-9.
Fuchs E, Tumbar T, Guasch G. Socializing with the neighbors: stem cells and their niche. Cell 2004; 116: 769-78.
Gomez-Lechon M, Jover R, Donato T, et al. Long-term expression of differentiated functions in hepatocytes cultured in three-dimensional collagen matrix. J Cell Physiol 1998; 77: 553-62.
Fischbach C, Chen R, Matsumoto T, et al. Egineering tumors with 3D scaffolds. Nat Methods 2007; 4: 855-60.
Gilbert PM, Havenstrite KL, Magnusson KE, et al. Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture. Science 2010; 329: 1078-81.
Engler A, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell 2006; 126: 677-89.
Hamburger A, Salmon SE. Primary bioassay of human tumor stem cells. Science 1977; 197: 461-3.
Mazzoleni G, Di Lorenzo D, Steimberg N. Modelling tissue in 3D: the next future of plarmaco-toxicology and food research? Genes Nutr 2009; 4: 13-22.
Vinci M, Gowan S, Boxall F, 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.
Vinci M, Box C, Eccles SA. Three-dimensional (3D) tumor spheroid invasion assay. J Vis Exp 2015; 99: e52686.
Chen S, Chang Y, Nieh S, et al. Nonadhesive culture system as a model of rapid sphere formation with cancer stem cell properties. PLoS One 2012; 7: e31864.
Griffith LG, Swartz MA. Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 2006; 7: 211-24.
Cawkill D, Eaglestone SS. Evolution of cell-based reagent provision. Drug Discov Today 2007; 12: 820-5.
Lee J, Cuddihy MJ, Kotov NA. Three-dimensional cell culture matrices: state of the art. Tissue Eng Part B Rev 2008; 14: 61-86.
Benya PD, Shaffer JD. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 1982; 30: 215-24.
Pradhan-Bhatt S, Harrington DA, Duncan RL, et al. A novel in vivo model for evaluating functional restoration of a tissue-engineered salivary gland. Laryngoscope 2014; 124: 456-61.
Pradhan S, Liu C, Zhang C, Jia X, Farach-Carson MC, Witt RL. Lumen formation in three-dimensional cultures of salivary acinar cells. Otolaryngol Head Neck Surg 2010; 142: 191-5.
Pradhan-Bhatt S, Harrington DA, Duncan RL, Jia X, Witt RL, Farach-Carson MC. Implantable three-dimensional salivary spheroid assemblies demonstrate fluid and protein secretory responses to neurotransmitters. Tissue Eng Part A 2013; 19: 1610-20.
Ghosh S, Spagnoli GC, Martin I, et al. Three-dimensional culture of melanoma cells profoundly affects gene expression profile: a high density oligonucleotide array study. J Cell Physiol 2005; 204: 522-31.
Berthiaume F, Moghe PV, Toner M, Yarmush ML. Effect of extracellular matrix topology on cell structure, function, and physiological responsiveness: hepatocytes cultured in a sandwich configuration. FASEB J 1996; 10: 1471-84.
Semino CE, Merok JR, Crane GG, Panagiotakos G, Zhang S. Functional differentiation of hepatocyte-like spheroid structures from putative liver progenitor cells in three-dimensional peptide scaffolds. Differentiation 2003; 71: 262-70.
Powers MJ, Janigian DM, Wack KE, Baker CS, Beer Stolz D, Griffith LG. Functional behavior of primary rat liver cells in a three-dimensional perfused microarray bioreactor. Tissue Eng 2002; 8: 499-513.
Frieboes HB, Smith BR, Chuang Y, et al. An integrated computational/experimental model of tumor invasion. Cancer Res 2006; 66: 1597-604.
Marushima H, Shibata S, Asakura T, et al. Three-dimensional culture promotes reconstitution of the tumor-specific hypoxic microenvironment under TGFβ stimulation. Int J Oncol 2011; 39: 1327-36.
Krishnamurthy S, Nör JE. Orosphere assay: a method for propagation of head and neck cancer stem cells. Head Neck 2013; 35: 1015-21.
Campos MS, Neiva KG, Meyers KA, Krishnamurthy S, Nör JE. Endothelial derived factors inhibit anoikis of head and neck cancer stem cells. Oral Oncol 2012; 48: 26-32.
Lo WL, Yu CC, Chiou GY, et al. MicroRNA-200c attenuates tumour growth and metastasis of presumptive head and neck squamous cell carcinoma stem cells. J Pathol 2011; 223: 482-95.
Hsieh C, Chen Y, Huang S, Wang H, Wu M. The effect of primary cancer cell culture models on the results of drug chemosensitivity assays: the application of perfusion microbioreactor system as cell culture vessel. Biomed Res Int 2015; 2015: 470283.
Bulysheva AA, Bowlin GL, Petrova SP, Yeudall WA. Enhanced chreemosistance of squamous carcinoma cells grown in 3D cryogenic electrospun scaffolds. Biomed Mater 2013; 8: 055009.
Storch K, Eke I, Borgmann K, et al. Three-dimensional cell growth confers radioresistance by chromatin density modification. Cancer Res 2010; 70: 3925-34.
Hehlgans S, Lange I, Eke I, Cordes N. 3D cell cultures of human head and neck squamous cell carcinoma cells are radiosensitized by the focal adhesion kinase inhibitor TAE226. Radiother Oncol 2009; 83: 371-8.
Eke I, Cordes N. Dual targeting of EGFR and focal adhesion kinase in 3D grown HNSCC cell cultures. Radiother Oncol 2011; 99: 279-86.
Luo Z, Ye T, Ma Y, Gill HS, Nitin N. Microprecision delivery of oligonucleotides in a 3D tissue model and its characterization using optical imaging. Mol Pharm 2013; 10: 2868-79.
Khetan S, Burdick JA. Patterning network structure to spatially control cellular remodeling and stem cell fate within 3-dimensional hydrogels. Biomaterials 2010; 31: 8228-34.
Weiswald L, Bellet D, Dangles-Marie V. Spherical cancer models in tumor biology. Neoplasia 2015; 17: 1-15.
Li Q, Chen C, Kapadia A, et al. 3D models of epithelial-mesenchymal transistion in breast cancer metastasis. J Biomol Screen 2011; 16: 141-54.
Heppner GH. Tumor heterogeneity. Cancer Res 1984; 44: 2259-65.
Paschos N, Brown WE, Eswaramoorthy R, Hu J, Athanasiou K. Advances in tissue engineering through stem cell-based co-cultures. J Tissue Eng Regen Med 2014; 9: 488-503.
Lawrance TS, Beers WH, Gilula NB. Transmission of hormonal stimulation by cell-to-cell communication. Nature 1978; 272: 501-6.
Hendriks J, Riesle J, van Blitterswijk CA. Co-culture in cartilage tissue engineering. J Tissue Eng Regen Med 2007; 1: 170-8.
Bian L, Zhai DY, Mauck RL, Burdick JA. Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage. Tissue Eng A 2011; 17: 1137-45.
Guan X, Delo DM, Atala A, Soker S. In vitro cardiomyogenic potential of human amniotic fluid stem cells. J Tissue Eng Regen Med 2011; 5: 220-8.
Vats A, Bielby RC, Tolley N, et al. Chondrogenic differentiation of human embryonic stem cells: the effect of the micro-evironment. Tissue Eng 2006; 12: 1687-97.
Traphagen S, Titushkin I, Sun S, Wary KK, Cho M. Endothielial invasive response in a co-culture model with physically-induced osteodifferentiation. J Tissue Eng Regen Med 2013; 7: 621-30.
Ou DB, He Y, Chen R, et al. Three-dimensional co-culture facilitates the differentiation of embryonic stem cells into mature cardiomyocytes. J Cell Biochem 2011; 112: 3555-62.
Kawada H, Ando K, Tsuji T, et al. Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system. Exp Hematol 1999; 27: 904-15.
Yamamoto Y, Mochida J, Sakai D, et al. Upregulation of the viability of nucleus pulposus cells by bone marrow-derived stromal cells: significance of direct cell-to-cell contact in coculture system. Spine 2004; 29: 1508-14.
Liu Y, Chan-Park MB. A biomimetic hydrogel based on methacrylated dextran-graft-lysine and gelatin for 3D smooth muscle cell culture. Biomaterials 2010; 31: 1158-70.
Goers L, Freemont P, Polizzi KM. Co-culture systems and technologies: taking synthetic biology to the next level. J R Soc Interface 2014; 11: 20140065.
Polson AG, Fuji RN. The successes and limitations of preclinical studies in predicting the pharmacodynamics and safety of cell-surface-targeted biological agents in patients. Br J Pharmacol 2012; 166: 1600-2.
Stasiak P, Sznitowska M. Zastosowanie hodowli komórkowych w badaniach biofarmaceutycznych. Farm Pol 2010; 66: 228-34.
Sodunke TR, Turner KK, Caldwell SA, McBride KW, Reginato MJ, Noh HM. Micropatterns of Matrigel for three-dimensional epithelial cultures. Biomaterials 2007; 28: 4006-16.
Friedrich J, Seidel C, Ebner R, Kunz-Schughart LA. Spheroid-based drug screen: considerations and practical approach. Nat Protoc 2009; 4: 309-24.
Kleinman HK, Martin GR. Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol 2005; 15: 378-86.
Dawson E, Mapili G, Erickson K, Tagvi S, Roy K. Biomaterials for stem cell differentiation. Adv Drug Deliv Rev 2008; 60: 215-28.
Kim JB. Three-dimensional tissue culture models in cancer biology. Semin Cancer Biol 2005; 15: 365-77.
Xu K, Buchsbaum RJ. Isolation of mammary epithelial cells from three-dimensional mixed-cell spheroid co-culture. J Vis Exp 2012; 62: pii: 3760.
Fridman R, Kibbey MC, Royce LS, et al. Enhanced tumor growth of both primary and established human and murine tumor cells in athymic mice after coinjection with matrigel. JNCI J Natl Cancer Inst 1991; 83: 769-74.
Jastrzebska K, Kucharczyk K, Florczak A, Dondajewska E, Mackiewicz A, Dams-Kozlowska H. Silk as an innovative biomaterial for cancer therapy. Rep Pract Oncol Radiother 2014; 20: 87-98.
Glicklis R, Shapiro L, Agbaria R, Merchuk JC, Cohen S. Hepatocyte behavior within three-dimensional porous alginate scaffolds. Biotechnol Bioeng 2000; 67: 344-53.
Tan W, Krishnaraj R, Desai TA. Evaluation of nanostructured composite collagen-chitosan matrices for tissue engineering. Tissue Eng 2001; 7: 203-10.
Sourla A, Doillon C, Koutsilieris M. Three-dimensional type I collagen gel system containing MG-63 osteoblasts-like cells as a model for studying local bone reaction caused by metastatic cancer cells. Anticancer Res 1996; 16: 2773-80.
Justice BA, Badr NA, Felder RA. 3D cell culture opens new dimension in cell-based assays. Drug Discov Today 2009; 14: 102-7.
Cheng N, Estes BT, Awad H, Guilak F. Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials 2004; 25: 3211-22.
Stevens MM, George JH. Exploring and engineering the cell surface interface. Science 2005; 310: 1135-8.
Curtis A, Wilkinson C. New depths in cell behaviour: reactions of cells to nanotopography. Biochem Soc Symp 1999; 65: 15-26.
Haycock JW. 3D cell culture: a review of current approaches and techniques. Methods Mol Biol 2011; 695: 1-15.
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