Three‑dimensional cell culture: A powerful tool in tumor research and drug discovery (Review)

Lv,Donglai; Hu,Zongtao; Lu,Lin; Lu,Husheng; Xu,Xiuli

doi:10.3892/ol.2017.7134

Three‑dimensional cell culture: A powerful tool in tumor research and drug discovery (Review)

Authors:
- Donglai Lv
- Zongtao Hu
- Lin Lu
- Husheng Lu
- Xiuli Xu
View Affiliations

Affiliations: Department of Clinical Oncology, The 105 Hospital of The People's Liberation Army, Hefei, Anhui 230031, P.R. China
Published online on: October 3, 2017 https://doi.org/10.3892/ol.2017.7134
Pages: 6999-7010

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

In previous years, three‑dimensional (3D) cell culture technology has become a focus of research in tumor cell biology, using a variety of methods and materials to mimic the in vivo microenvironment of cultured tumor cells ex vivo. These 3D tumor cells have demonstrated numerous different characteristics compared with traditional two‑dimensional (2D) culture. 3D cell culture provides a useful platform for further identifying the biological characteristics of tumor cells, particularly in the drug sensitivity area of the key points of translational medicine. It promises to be a bridge between traditional 2D culture and animal experiments, and is of great importance for further research in the field of tumor biology. In the present review, previous 3D cell culture applications, focusing on anti‑tumor drug susceptibility testing, are summarized.

View Figures

View References

1	Abbott A: Cell culture: Biology's new dimension. Nature. 424:870–872. 2003. View Article : Google Scholar : PubMed/NCBI
2	Weigelt B, Ghajar CM and Bissell MJ: The need for complex 3D culture models to unravel novel pathways and identify accurate biomarkers in breast cancer. Adv Drug Deliv Rev. 69–70:42–51. 2014. View Article : Google Scholar
3	Jacks T and Weinberg RA: Taking the study of cancer cell survival to a new dimension. Cell. 111:923–925. 2002. View Article : Google Scholar : PubMed/NCBI
4	Breslin S and O'Driscoll L: Three-dimensional cell culture: The missing link in drug discovery. Drug Discov Today. 18:240–249. 2013. View Article : Google Scholar : PubMed/NCBI
5	Griffith LG and Swartz MA: Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol. 7:211–224. 2006. View Article : Google Scholar : PubMed/NCBI
6	Chen L, Xiao Z, Meng Y, Zhao Y, Han J, Su G, Chen B and Dai J: The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs. Biomaterials. 33:1437–1444. 2012. View Article : Google Scholar : PubMed/NCBI
7	Dhiman HK, Ray AR and Panda AK: Three-dimensional chitosan scaffold-based MCF-7 cell culture for the determination of the cytotoxicity of tamoxifen. Biomaterials. 26:979–986. 2005. View Article : Google Scholar : PubMed/NCBI
8	Fong EL, Lamhamedi-Cherradi SE, Burdett E, Ramamoorthy V, Lazar AJ, Kasper FK, Farach-Carson MC, Vishwamitra D, Demicco EG, Menegaz BA, et al: Modeling Ewing sarcoma tumors in vitro with 3D scaffolds. Proc Natl Acad Sci USA. 110:pp. 6500–6505. 2013, View Article : Google Scholar : PubMed/NCBI
9	Wästfelt M, Fadeel B and Henter JI: A journey of hope: Lessons learned from studies on rare diseases and orphan drugs. J Intern Med. 260:1–10. 2006. View Article : Google Scholar : PubMed/NCBI
10	Hutmacher DW: Biomaterials offer cancer research the third dimension. Nat Mater. 9:90–93. 2010. View Article : Google Scholar : PubMed/NCBI
11	Shoemaker RH: The NCI60 human tumour cell line anticancer drug screen. Nat Rev Cancer. 6:813–823. 2006. View Article : Google Scholar : PubMed/NCBI
12	Desrochers TM, Palma E and Kaplan DL: Tissue-engineered kidney disease models. Adv Drug Deliv Rev. 69–70:67–80. 2014. View Article : Google Scholar
13	Cree IA, Glaysher S and Harvey AL: Efficacy of anti-cancer agents in cell lines versus human primary tumour tissue. Curr Opin Pharmacol. 10:375–379. 2010. View Article : Google Scholar : PubMed/NCBI
14	Uchida Y, Tanaka S, Aihara A, Adikrisna R, Yoshitake K, Matsumura S, Mitsunori Y, Murakata A, Noguchi N, Irie T, et al: Analogy between sphere forming ability and stemness of human hepatoma cells. Oncol Rep. 24:1147–1151. 2010.PubMed/NCBI
15	Stevens JL and Baker TK: The future of drug safety testing: Expanding the view and narrowing the focus. Drug Discov Today. 14:162–167. 2009. View Article : Google Scholar : PubMed/NCBI
16	Huh D, Matthews BD, Mammoto A, Montoya-Zavala M, Hsin HY and Ingber DE: Reconstituting organ-level lung functions on a chip. Science. 328:1662–1668. 2010. View Article : Google Scholar : PubMed/NCBI
17	Russell WMS and Burch RL: The principles of humane experimental technique. Methuen; London: 1959
18	Talukdar S and Kundu SC: A non-mulberry silk fibroin protein based 3d in vitro tumor model for evaluation of anticancer drug activity. Adv Funct Mat. 22:4778–4788. 2012. View Article : Google Scholar
19	Dunne LW, Huang Z, Meng W, Fan X, Zhang N, Zhang Q and An Z: Human decellularized adipose tissue scaffold as a model for breast cancer cell growth and drug treatments. Biomaterials. 35:4940–4949. 2014. View Article : Google Scholar : PubMed/NCBI
20	Maguire SL, Peck B, Wai PT, Campbell J, Barker H, Gulati A, Daley F, Vyse S, Huang P, Lord CJ, et al: Three-dimensional modelling identifies novel genetic dependencies associated with breast cancer progression in the isogenic MCF10 model. J Pathol. 240:315–328. 2016. View Article : Google Scholar : PubMed/NCBI
21	Sha H, Zou Z, Xin K, Bian X, Cai X, Lu W, Chen J, Chen G, Huang L, Blair AM, et al: Tumor-penetrating peptide fused EGFR single-domain antibody enhances cancer drug penetration into 3D multicellular spheroids and facilitates effective gastric cancer therapy. J Control Release. 200:188–200. 2015. View Article : Google Scholar : PubMed/NCBI
22	Kundu B, Saha P, Datta K and Kundu SC: A silk fibroin based hepatocarcinoma model and the assessment of the drug response in hyaluronan-binding protein 1 overexpressed HepG2 cells. Biomaterials. 34:9462–9474. 2013. View Article : Google Scholar : PubMed/NCBI
23	Xu Z, Gao Y, Hao Y, Li E, Wang Y, Zhang J, Wang W, Gao Z and Wang Q: Application of a microfluidic chip-based 3D co-culture to test drug sensitivity for individualized treatment of lung cancer. Biomaterials. 34:4109–4117. 2013. View Article : Google Scholar : PubMed/NCBI
24	Simon KA, Mosadegh B, Minn KT, Lockett MR, Mohammady MR, Boucher DM, Hall AB, Hillier SM, Udagawa T, Eustace BK and Whitesides GM: Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system. Biomaterials. 95:47–59. 2016. View Article : Google Scholar : PubMed/NCBI
25	Stratmann AT, Fecher D, Wangorsch G, Göttlich C, Walles T, Walles H, Dandekar T, Dandekar G and Nietzer SL: Establishment of a human 3D lung cancer model based on a biological tissue matrix combined with a Boolean in silico model. Mol Oncol. 8:351–365. 2014. View Article : Google Scholar : PubMed/NCBI
26	Lee JM, Mhawech-Fauceglia P, Lee N, Parsanian LC, Lin YG, Gayther SA and Lawrenson K: A three-dimensional microenvironment alters protein expression and chemosensitivity of epithelial ovarian cancer cells in vitro. Lab Invest. 93:528–542. 2013. View Article : Google Scholar : PubMed/NCBI
27	Shin CS, Kwak B, Han B and Park K: Development of an in vitro 3D tumor model to study therapeutic efficiency of an anticancer drug. Mol Pharm. 10:2167–2175. 2013. View Article : Google Scholar : PubMed/NCBI
28	Loessner D, Rizzi SC, Stok KS, Fuehrmann T, Hollier B, Magdolen V, Hutmacher DW and Clements JA: A bioengineered 3D ovarian cancer model for the assessment of peptidase-mediated enhancement of spheroid growth and intraperitoneal spread. Biomaterials. 34:7389–7400. 2013. View Article : Google Scholar : PubMed/NCBI
29	Yang Z and Zhao X: A 3D model of ovarian cancer cell lines on peptide nanofiber scaffold to explore the cell-scaffold interaction and chemotherapeutic resistance of anticancer drugs. Int J Nanomedicine. 6:303–310. 2011. View Article : Google Scholar : PubMed/NCBI
30	Fitzgerald KA, Guo J, Tierney EG, Curtin CM, Malhotra M, Darcy R, O'Brien FJ and O'Driscoll CM: The use of collagen-based scaffolds to simulate prostate cancer bone metastases with potential for evaluating delivery of nanoparticulate gene therapeutics. Biomaterials. 66:53–66. 2015. View Article : Google Scholar : PubMed/NCBI
31	Xu X, Sabanayagam CR, Harrington DA, Farach-Carson MC and Jia X: A hydrogel-based tumor model for the evaluation of nanoparticle-based cancer therapeutics. Biomaterials. 35:3319–3330. 2014. View Article : Google Scholar : PubMed/NCBI
32	Lv D, Yu SC, Ping YF, Wu H, Zhao X, Zhang H, Cui Y, Chen B, Zhang X, Dai J, et al: A three-dimensional collagen scaffold cell culture system for screening anti-glioma therapeutics. Oncotarget. 7:56904–56914. 2016. View Article : Google Scholar : PubMed/NCBI
33	Ma NK, Lim JK, Leong MF, Sandanaraj E, Ang BT, Tang C and Wan AC: Collaboration of 3D context and extracellular matrix in the development of glioma stemness in a 3D model. Biomaterials. 78:62–73. 2016. View Article : Google Scholar : PubMed/NCBI
34	Pedron S, Becka E and Harley BA: Regulation of glioma cell phenotype in 3D matrices by hyaluronic acid. Biomaterials. 34:7408–7417. 2013. View Article : Google Scholar : PubMed/NCBI
35	Munson JM, Bellamkonda RV and Swartz MA: Interstitial flow in a 3D microenvironment increases glioma invasion by a CXCR4-dependent mechanism. Cancer Res. 73:1536–1546. 2013. View Article : Google Scholar : PubMed/NCBI
36	Fennema E, Rivron N, Rouwkema J, van Blitterswijk C and de Boer J: Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol. 31:108–115. 2013. View Article : Google Scholar : PubMed/NCBI
37	Friedrich J, Seidel C, Ebner R and Kunz-Schughart LA: Spheroid-based drug screen: Considerations and practical approach. Nat Protoc. 4:309–324. 2009. View Article : Google Scholar : PubMed/NCBI
38	Reynolds BA and Weiss S: Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science. 255:1707–1710. 1992. View Article : Google Scholar : PubMed/NCBI
39	Yu SC, Ping YF, Yi L, Zhou ZH, Chen JH, Yao XH, Gao L, Wang JM and Bian XW: Isolation and characterization of cancer stem cells from a human glioblastoma cell line U87. Cancer Lett. 265:124–134. 2008. View Article : Google Scholar : PubMed/NCBI
40	Yuhas JM, Li AP, Martinez AO and Ladman AJ: A simplified method for production and growth of multicellular tumor spheroids. Cancer Res. 37:3639–3643. 1977.PubMed/NCBI
41	Lawlor ER, Scheel C, Irving J and Sorensen PH: Anchorage-independent multi-cellular spheroids as an in vitro model of growth signaling in Ewing tumors. Oncogene. 21:307–318. 2002. View Article : Google Scholar : PubMed/NCBI
42	Lin RZ and Chang HY: Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol J. 3:1172–1184. 2008. View Article : Google Scholar : PubMed/NCBI
43	Pang R, Law WL, Chu AC, Poon JT, Lam CS, Chow AK, Ng L, Cheung LW, Lan XR, Lan HY, et al: A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. Cell Stem Cell. 6:603–615. 2010. View Article : Google Scholar : PubMed/NCBI
44	Ivascu A and Kubbies M: Rapid generation of single-tumor spheroids for high-throughput cell function and toxicity analysis. J Biomol Screen. 11:922–932. 2006. View Article : Google Scholar : PubMed/NCBI
45	Louis SA, Rietze RL, Deleyrolle L, Wagey RE, Thomas TE, Eaves AC and Reynolds BA: Enumeration of neural stem and progenitor cells in the neural colony-forming cell assay. Stem Cells. 26:988–996. 2008. View Article : Google Scholar : PubMed/NCBI
46	Ng KW, Leong DT and Hutmacher DW: The challenge to measure cell proliferation in two and three dimensions. Tissue Eng. 11:182–191. 2005. View Article : Google Scholar : PubMed/NCBI
47	Kelm JM, Timmins NE, Brown CJ, Fussenegger M and Nielsen LK: Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng. 83:173–180. 2003. View Article : Google Scholar : PubMed/NCBI
48	Tung YC, Hsiao AY, Allen SG, Torisawa YS, Ho M and Takayama S: High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. Analyst. 136:473–478. 2011. View Article : Google Scholar : PubMed/NCBI
49	Zanetta M, Quirici N, Demarosi F, Tanzi MC, Rimondini L and Farè S: Ability of polyurethane foams to support cell proliferation and the differentiation of MSCs into osteoblasts. Acta Biomater. 5:1126–1136. 2009. View Article : Google Scholar : PubMed/NCBI
50	Souza GR, Molina JR, Raphael RM, Ozawa MG, Stark DJ, Levin CS, Bronk LF, Ananta JS, Mandelin J, Georgescu MM, et al: Three-dimensional tissue culture based on magnetic cell levitation. Nat Nanotechnol. 5:291–296. 2010. View Article : Google Scholar : PubMed/NCBI
51	Su J, Zhang L, Zhang W, Choi DS, Wen J, Jiang B, Chang CC and Zhou X: Targeting the biophysical properties of the myeloma initiating cell niches: A pharmaceutical synergism analysis using multi-scale agent-based modeling. PLoS One. 9:e850592014. View Article : Google Scholar : PubMed/NCBI
52	Durand RE and Sutherland RM: Effects of intercellular contact on repair of radiation damage. Exp Cell Res. 71:75–80. 1972. View Article : Google Scholar : PubMed/NCBI
53	Goodwin TJ, Prewett TL, Wolf DA and Spaulding GF: Reduced shear stress: A major component in the ability of mammalian tissues to form three-dimensional assemblies in simulated microgravity. J Cell Biochem. 51:301–311. 1993. View Article : Google Scholar : PubMed/NCBI
54	Chen X, Xu H, Wan C, McCaigue M and Li G: Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells. Stem Cells. 24:2052–2059. 2006. View Article : Google Scholar : PubMed/NCBI
55	Luni C, Feldman HC, Pozzobon M, De Coppi P, Meinhart CD and Elvassore N: Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture. Biomicrofluidics. 4:pii: 034105. 2010. View Article : Google Scholar : PubMed/NCBI
56	Chang TT and Hughes-Fulford M: Monolayer and spheroid culture of human liver hepatocellular carcinoma cell line cells demonstrate distinct global gene expression patterns and functional phenotypes. Tissue Eng Part A. 15:559–567. 2009. View Article : Google Scholar : PubMed/NCBI
57	Redden RA and Doolin EJ: Microgravity assay of neuroblastoma: In vitro aggregation kinetics and organoid morphology correlate with MYCN expression. In Vitro Cell Dev Biol Anim. 47:312–317. 2011. View Article : Google Scholar : PubMed/NCBI
58	Kaur P, Ward B, Saha B, Young L, Groshen S, Techy G, Lu Y, Atkinson R, Taylor CR, Ingram M and Imam SA: Human breast cancer histoid: An in vitro 3-dimensional co-culture model that mimics breast cancer tissue. J Histochem Cytochem. 59:1087–1100. 2011. View Article : Google Scholar : PubMed/NCBI
59	Marrero B, Messina JL and Heller R: Generation of a tumor spheroid in a microgravity environment as a 3D model of melanoma. In Vitro Cell Dev Biol Anim. 45:523–534. 2009. View Article : Google Scholar : PubMed/NCBI
60	Zhang Z and Nagrath S: Microfluidics and cancer: Are we there yet? Biomed Microdevices. 15:595–609. 2013. View Article : Google Scholar : PubMed/NCBI
61	Mehta G, Hsiao AY, Ingram M, Luker GD and Takayama S: Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release. 164:192–204. 2012. View Article : Google Scholar : PubMed/NCBI
62	Gill BJ and West JL: Modeling the tumor extracellular matrix: Tissue engineering tools repurposed towards new frontiers in cancer biology. J Biomech. 47:1969–1978. 2014. View Article : Google Scholar : PubMed/NCBI
63	Loessner D, Stok KS, Lutolf MP, Hutmacher DW, Clements JA and Rizzi SC: Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. Biomaterials. 31:8494–8506. 2010. View Article : Google Scholar : PubMed/NCBI
64	Nirmalanandhan VS, Duren A, Hendricks P, Vielhauer G and Sittampalam GS: Activity of anticancer agents in a three-dimensional cell culture model. Assay Drug Dev Technol. 8:581–590. 2010. View Article : Google Scholar : PubMed/NCBI
65	Freeman AE and Hoffman RM: In vivo-like growth of human tumors in vitro. Proc Natl Acad Sci USA. 83:pp. 2694–2698. 1986, View Article : Google Scholar : PubMed/NCBI
66	Takamura Y, Kobayashi H, Taguchi T, Motomura K, Inaji H and Noguchi S: Prediction of chemotherapeutic response by collagen gel droplet embedded culture-drug sensitivity test in human breast cancers. Int J Cancer. 98:450–455. 2002. View Article : Google Scholar : PubMed/NCBI
67	Yang J, Richards J, Bowman P, Guzman R, Enami J, McCormick K, Hamamoto S, Pitelka D and Nandi S: Sustained growth and three-dimensional organization of primary mammary tumor epithelial cells embedded in collagen gels. Proc Natl Acad Sci USA. 76:pp. 3401–3405. 1979, View Article : Google Scholar : PubMed/NCBI
68	Rowehl RA, Burke S, Bialkowska AB, Pettet DW III, Rowehl L, Li E, Antoniou E, Zhang Y, Bergamaschi R, Shroyer KR, et al: Establishment of highly tumorigenic human colorectal cancer cell line (CR4) with properties of putative cancer stem cells. PLoS One. 9:e990912014. View Article : Google Scholar : PubMed/NCBI
69	Yip D and Cho CH: A multicellular 3D heterospheroid model of liver tumor and stromal cells in collagen gel for anti-cancer drug testing. Biochem Biophys Res Commun. 433:327–332. 2013. View Article : Google Scholar : PubMed/NCBI
70	Rao SS, Dejesus J, Short AR, Otero JJ, Sarkar A and Winter JO: Glioblastoma behaviors in three-dimensional collagen-hyaluronan composite hydrogels. ACS Appl Mater Interfaces. 5:9276–9284. 2013. View Article : Google Scholar : PubMed/NCBI
71	Dvir-Ginzberg M, Gamlieli-Bonshtein I, Agbaria R and Cohen S: Liver tissue engineering within alginate scaffolds: Effects of cell-seeding density on hepatocyte viability, morphology, and function. Tissue Eng. 9:757–766. 2003. View Article : Google Scholar : PubMed/NCBI
72	Zhang X, Wang W, Yu W, Xie Y, Zhang Y and Ma X: Development of an in vitro multicellular tumor spheroid model using microencapsulation and its application in anticancer drug screening and testing. Biotechnol Prog. 21:1289–1296. 2005. View Article : Google Scholar : PubMed/NCBI
73	Xu XX, Liu C, Liu Y, Yang L, Li N, Guo X, Sun GW and Ma XJ: Enrichment of cancer stem cell-like cells by culture in alginate gel beads. J Biotechnol. 177:1–12. 2014. View Article : Google Scholar : PubMed/NCBI
74	Kleinman HK and Martin GR: Matrigel: Basement membrane matrix with biological activity. Semin Cancer Biol. 15:378–386. 2005. View Article : Google Scholar : PubMed/NCBI
75	Wang F, Weaver VM, Petersen OW, Larabell CA, Dedhar S, Briand P, Lupu R and Bissell MJ: Reciprocal interactions between beta1-integrin and epidermal growth factor receptor in three-dimensional basement membrane breast cultures: A different perspective in epithelial biology. Proc Natl Acad Sci USA. 95:pp. 14821–14826. 1998, View Article : Google Scholar : PubMed/NCBI
76	Ampuja M, Jokimäki R, Juuti-Uusitalo K, Rodriguez-Martinez A, Alarmo EL and Kallioniemi A: BMP4 inhibits the proliferation of breast cancer cells and induces an MMP-dependent migratory phenotype in MDA-MB-231 cells in 3D environment. BMC Cancer. 13:4292013. View Article : Google Scholar : PubMed/NCBI
77	Christensen M, Najy AJ, Snyder M, Movilla LS and Kim HR: A critical role of the PTEN/PDGF signaling network for the regulation of radiosensitivity in adenocarcinoma of the prostate. Int J Radiat Oncol Biol Phys. 88:151–158. 2014. View Article : Google Scholar : PubMed/NCBI
78	Valyi-Nagy K, Kormos B, Ali M, Shukla D and Valyi-Nagy T: Stem cell marker CD271 is expressed by vasculogenic mimicry-forming uveal melanoma cells in three-dimensional cultures. Mol Vis. 18:588–592. 2012.PubMed/NCBI
79	Lombardo Y, Filipović A, Molyneux G, Periyasamy M, Giamas G, Hu Y, Trivedi PS, Wang J, Yagüe E, Michel L and Coombes RC: Nicastrin regulates breast cancer stem cell properties and tumor growth in vitro and in vivo. Proc Natl Acad Sci USA. 109:pp. 16558–16563. 2012, View Article : Google Scholar : PubMed/NCBI
80	Sodunke TR, Turner KK, Caldwell SA, McBride KW, Reginato MJ and Noh HM: Micropatterns of Matrigel for three-dimensional epithelial cultures. Biomaterials. 28:4006–4016. 2007. View Article : Google Scholar : PubMed/NCBI
81	Carletti E, Motta A and Migliaresi C: Scaffolds for tissue engineering and 3D cell culture. Methods Mol Biol. 695:17–39. 2011. View Article : Google Scholar : PubMed/NCBI
82	Liu X, Holzwarth JM and Ma PX: Functionalized synthetic biodegradable polymer scaffolds for tissue engineering. Macromol Biosci. 12:911–919. 2012. View Article : Google Scholar : PubMed/NCBI
83	Feng S, Duan X, Lo PK, Liu S, Liu X, Chen H and Wang Q: Expansion of breast cancer stem cells with fibrous scaffolds. Integr Biol (Camb). 5:768–777. 2013. View Article : Google Scholar : PubMed/NCBI
84	Chen J, Wang J, Zhang Y, Chen D, Yang C, Kai C, Wang X, Shi F and Dou J: Observation of ovarian cancer stem cell behavior and investigation of potential mechanisms of drug resistance in three-dimensional cell culture. J Biosci Bioeng. 118:214–222. 2014. View Article : Google Scholar : PubMed/NCBI
85	Kievit FM, Florczyk SJ, Leung MC, Wang K, Wu JD, Silber JR, Ellenbogen RG, Lee JS and Zhang M: Proliferation and enrichment of CD133(+) glioblastoma cancer stem cells on 3D chitosan-alginate scaffolds. Biomaterials. 35:9137–9143. 2014. View Article : Google Scholar : PubMed/NCBI
86	Amann A, Zwierzina M, Gamerith G, Bitsche M, Huber JM, Vogel GF, Blumer M, Koeck S, Pechriggl EJ, Kelm JM, et al: Development of an innovative 3D cell culture system to study tumour-stroma interactions in non-small cell lung cancer cells. PLoS One. 9:e925112014. View Article : Google Scholar : PubMed/NCBI
87	Hoque MT, Windus LC, Lovitt CJ and Avery VM: PCaAnalyser: A 2D-image analysis based module for effective determination of prostate cancer progression in 3D culture. PLoS One. 8:e798652013. View Article : Google Scholar : PubMed/NCBI
88	Ciurea ME, Georgescu AM, Purcaru SO, Artene SA, Emami GH, Boldeanu MV, Tache DE and Dricu A: Cancer stem cells: Biological functions and therapeutically targeting. Int J Mol Sci. 15:8169–8185. 2014. View Article : Google Scholar : PubMed/NCBI