3D tumour models: novel in vitro approaches to cancer studies

Review

Abstract

3D in vitro models have been used in cancer research as a compromise between 2-dimensional cultures of isolated cancer cells and the manufactured complexity of xenografts of human cancers in immunocompromised animal hosts. 3D models can be tailored to be biomimetic and accurately recapitulate the native in vivo scenario in which they are found. These 3D in vitro models provide an important alternative to both complex in vivo whole organism approaches, and 2D culture with its spatial limitations. Approaches to create more biomimetic 3D models of cancer include, but are not limited to, (i) providing the appropriate matrix components in a 3D configuration found in vivo, (ii) co-culturing cancer cells, endothelial cells and other associated cells in a spatially relevant manner, (iii) monitoring and controlling hypoxia- to mimic levels found in native tumours and (iv) monitoring the release of angiogenic factors by cancer cells in response to hypoxia. This article aims to overview current 3D in vitro models of cancer and review strategies employed by researchers to tackle these aspects with special reference to recent promising developments, as well as the current limitations of 2D cultures and in vivo models. 3D in vitro models provide an important alternative to both complex in vivo whole organism approaches, and 2D culture with its spatial limitations. Here we review current strategies in the field of modelling cancer, with special reference to advances in complex 3D in vitro models.

Keywords

Biomimetic Tumour stroma 3D cancer models In vitro tumour models 

List of abbreviations

2D

Two-dimensional

3D

Three-dimensional

bFGF

Basic fibroblast growth factor

BME

Basement membrane extract

BSA

Bovine serum albumin

DOX

Doxorubicin

EC

Endothelial cell

ECM

Extracellular matrix

EGF

Epidermal growth factor

EHS

Engelbreth-Holm-Swarm

EOC

Human epithelial ovarian cancer

HA

Hyaluronan / hyaluronic acid

IL-8

Interleukin-8

lrECM

Laminin-rich extracellular matrix

MCS

Mesenchymal stem cells

MCTS

Multicellular tumour spheroid

MMP

Metalloproteinase

NOD

Non-obese diabetic

PBS

Phosphate buffered saline

PC

Plastic compression

PGA

Polyglycolide

PEG

Polyethylene glycol

PLA

Polylactide

PLG/PLGA

Poly(lactide-co-glycolide)

PVA

Poly(vinyl alcohol)

RGD

Arginine-glycine-aspartic acid

SCID

Severely compromised immunodeficient

VEGF

Vascular endothelial growth factor

Notes

Acknowledgements

Umber Cheema is a BBSRC David Phillips Fellow and is funded through this route.

References

  1. Abou Neel EA, Cheema U, Knowles JC, Brown RA, Nazhat SN (2006) Use of multiple unconfined compression for control of collagen gel scaffold density and mechanical properties. Soft Matter 2:986–992CrossRefGoogle Scholar
  2. Agrawal CM, Ray RB (2001) Biodegradable polymeric scaffolds for musculoskeletal tissue engineering. J Biomed Mater Res 55:141–150PubMedCrossRefGoogle Scholar
  3. Albihn A, Johnsen JI, Henriksson MA (2010) MYC in oncogenesis and as a target for cancer therapies. Adv Cancer Res 107:163–224PubMedCrossRefGoogle Scholar
  4. Amatangelo MD, Bassi DE, Klein-Szanto AJ, Cukierman E (2005) Stroma-derived three-dimensional matrices are necessary and sufficient to promote desmoplastic differentiation of normal fibroblasts. Am J Pathol 167:475–488PubMedCrossRefGoogle Scholar
  5. Boehm T, Folkman J, Browder T, O’Reilly MS (1997) Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390:404–407PubMedCrossRefGoogle Scholar
  6. Brown RA, Wiseman M, Chuo C-B, Cheema U, Nazhat SN (2005) Ultrarapid engineering of biomimetic biomaterials and tissues: fabrication of nano- and microstructures by plastic compression. Adv Funct Mater 15:1762–1770CrossRefGoogle Scholar
  7. Calmels TP, Mattot V, Wernert N, Vandenbunder B, Stehelin D (1995) Invasive tumors induce c-ets1 transcription factor expression in adjacent stroma. Biol Cell 84:53–61PubMedCrossRefGoogle Scholar
  8. Castello-Cros R, Khan DR, Simons J, Valianou M, Cukierman E (2009) Staged stromal extracellular 3D matrices differentially regulate breast cancer cell responses through PI3K and beta1-integrins. BMC Cancer 9:94PubMedCrossRefGoogle Scholar
  9. Cheema U, Yang SY, Mudera V, Goldspink GG, Brown RA (2003) 3-D in vitro model of early skeletal muscle development. Cell Motil Cytoskeleton 54:226–236PubMedCrossRefGoogle Scholar
  10. Cheema U, Brown RA, Alp B, Macrobert AJ (2008) Spatially defined oxygen gradients and vascular endothelial growth factor expression in an engineered 3D cell model. Cell Mol Life Sci 65:177–186PubMedCrossRefGoogle Scholar
  11. Cheema U, Alekseeva T, Abou-Neel EA, Brown RA (2010) Switching off angiogenic signalling: creating channelled constructs for adequate oxygen delivery in tissue engineered constructs. Eur Cell Mater 20:274–280PubMedGoogle Scholar
  12. Chen G, Sato T, Ushida T, Hirochika R, Shirasaki Y, Ochiai N, Tateishi T (2003) The use of a novel PLGA fiber/collagen composite web as a scaffold for engineering of articular cartilage tissue with adjustable thickness. J Biomed Mater Res A 67:1170–1180PubMedCrossRefGoogle Scholar
  13. Chen R, Khormaee S, Eccleston ME, Slater NK (2009) The role of hydrophobic amino acid grafts in the enhancement of membrane-disruptive activity of pH-responsive pseudo-peptides. Biomaterials 30:1954–1961PubMedCrossRefGoogle Scholar
  14. Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712PubMedCrossRefGoogle Scholar
  15. Cunliffe D, Pennadam S, Alexander C (2004) Synthetic and biological polymers—merging the interface. Eur Polym J 40:5–25CrossRefGoogle Scholar
  16. David L, Dulong V, Le CD, Chauzy C, Norris V, Delpech B, Lamacz M, Vannier JP (2004) Reticulated hyaluronan hydrogels: a model for examining cancer cell invasion in 3D. Matrix Biol 23:183–193PubMedCrossRefGoogle Scholar
  17. Dedhar S, Hannigan GE (1996) Integrin cytoplasmic interactions and bidirectional transmembrane signalling. Curr Opin Cell Biol 8:657–669PubMedCrossRefGoogle Scholar
  18. Eder JP Jr, Supko JG, Clark JW, Puchalski TA, Garcia-Carbonero R, Ryan DP, Shulman LN, Proper J, Kirvan M, Rattner B, Connors S, Keogan MT, Janicek MJ, Fogler WE, Schnipper L, Kinchla N, Sidor C, Phillips E, Folkman J, Kufe DW (2002) Phase I clinical trial of recombinant human endostatin administered as a short intravenous infusion repeated daily. J Clin Oncol 20:3772–3784PubMedCrossRefGoogle Scholar
  19. Elkas JC, Baldwin RL, Pegram M, Tseng Y, Slamon D, Karlan BY (2002) A human ovarian carcinoma murine xenograft model useful for preclinical trials. Gynecol Oncol 87:200–206PubMedCrossRefGoogle Scholar
  20. Fan BT, Lapluye G, Gavach C (1987) Potential study of basement membrane. Biochim Biophys Acta 900:183–190PubMedCrossRefGoogle Scholar
  21. Fischbach C, Chen R, Matsumoto T, Schmelzle T, Brugge JS, Polverini PJ, Mooney DJ (2007) Engineering tumors with 3D scaffolds. Nat Meth 4:855–860CrossRefGoogle Scholar
  22. Freyer JP, Sutherland RM (1980) Selective dissociation and characterization of cells from different regions of multicell tumor spheroids. Cancer Res 40:3956–3965PubMedGoogle Scholar
  23. Gurski LA, Jha AK, Zhang C, Jia X, Farach-Carson MC (2009) Hyaluronic acid-based hydrogels as 3D matrices for in vitro evaluation of chemotherapeutic drugs using poorly adherent prostate cancer cells. Biomaterials 30:6076–6085PubMedCrossRefGoogle Scholar
  24. Harris AL (2002) Hypoxia—a key regulatory factor in tumour growth. Nat Rev Cancer 2:38–47PubMedCrossRefGoogle Scholar
  25. Hemler ME, Mannion BA, Berditchevski F (1996) Association of TM4SF proteins with integrins: relevance to cancer. Biochim Biophys Acta 1287:67–71PubMedGoogle Scholar
  26. Ho VH, Slater NK, Chen R (2011) pH-responsive endosomolytic pseudo-peptides for drug delivery to multicellular spheroids tumour models. Biomaterials 32:2953–2958PubMedCrossRefGoogle Scholar
  27. Holliday DL, Brouilette KT, Markert A, Gordon LA, Jones JL (2009) Novel multicellular organotypic models of normal and malignant breast: tools for dissecting the role of the microenvironment in breast cancer progression. Breast Cancer Res 11:R3PubMedCrossRefGoogle Scholar
  28. Hutmacher DW (2000) Scaffolds in tissue engineering bone and cartilage. Biomaterials 21:2529–2543PubMedCrossRefGoogle Scholar
  29. Kenny PA, Lee GY, Myers CA, Neve RM, Semeiks JR, Spellman PT, Lorenz K, Lee EH, Barcellos-Hoff MH, Petersen OW, Gray JW, Bissell MJ (2007) The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression. Mol Oncol 1:84–96PubMedCrossRefGoogle Scholar
  30. Kim TH, Mount CW, Gombotz WR, Pun SH (2010) The delivery of doxorubicin to 3-D multicellular spheroids and tumors in a murine xenograft model using tumor-penetrating triblock polymeric micelles. Biomaterials 31:7386–7397PubMedCrossRefGoogle Scholar
  31. Kleinman HK, Martin GR (2005) Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol 15:378–386PubMedCrossRefGoogle Scholar
  32. Kramer RH, Bensch KG, Wong J (1986) Invasion of reconstituted basement membrane matrix by metastatic human tumor cells. Cancer Res 46:1980–1989PubMedGoogle Scholar
  33. Kunz-Schughart LA, Kreutz M, Knuechel R (1998) Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol 79:1–23PubMedCrossRefGoogle Scholar
  34. Lee GY, Kenny PA, Lee EH, Bissell MJ (2007) Three-dimensional culture models of normal and malignant breast epithelial cells. Nat Meth 4:359–365CrossRefGoogle Scholar
  35. Loessner D, Stok KS, Lutolf MP, Hutmacher DW, Clements JA, Rizzi SC (2010) Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. Biomaterials 31:8494–8506PubMedCrossRefGoogle Scholar
  36. Loudos G, Kagadis GC, Psimadas D (2010) Current status and future perspectives of in vivo small animal imaging using radiolabeled nanoparticles. Eur J RadiolGoogle Scholar
  37. Mandal BB, Kundu SC (2007) A novel method for dissolution and stabilization of non-mulberry silk gland protein fibroin using anionic surfactant sodium dodecyl sulfate. Biotechnol Bioeng 99:1482–1489CrossRefGoogle Scholar
  38. Monazzam A, Razifar P, Simonsson M, Qvarnstrom F, Josephsson R, Blomqvist C, Langstrom B, Bergstrom M (2006) Multicellular tumour spheroid as a model for evaluation of [18F]FDG as biomarker for breast cancer treatment monitoring. Cancer Cell Int 6:6PubMedCrossRefGoogle Scholar
  39. Monazzam A, Razifar P, Ide S, Rugaard JM, Josephsson R, Blomqvist C, Langstrom B, Bergstrom M (2009) Evaluation of the Hsp90 inhibitor NVP-AUY922 in multicellular tumour spheroids with respect to effects on growth and PET tracer uptake. Nucl Med Biol 36:335–342PubMedCrossRefGoogle Scholar
  40. Mueller-Klieser W (1997) Three-dimensional cell cultures: from molecular mechanisms to clinical applications. Am J Physiol 273:C1109–C1123PubMedGoogle Scholar
  41. Nilsson EE, Westfall SD, McDonald C, Lison T, Sadler-Riggleman I, Skinner MK (2002) An in vivo mouse reporter gene (human secreted alkaline phosphatase) model to monitor ovarian tumor growth and response to therapeutics. Cancer Chemother Pharmacol 49:93–100PubMedCrossRefGoogle Scholar
  42. Noskova V, Ahmadi S, Asander E, Casslen B (2009) Ovarian cancer cells stimulate uPA gene expression in fibroblastic stromal cells via multiple paracrine and autocrine mechanisms. Gynecol Oncol 115:121–126PubMedCrossRefGoogle Scholar
  43. Ong SM, Zhao Z, Arooz T, Zhao D, Zhang S, Du T, Wasser M, van ND, Yu H (2010) Engineering a scaffold-free 3D tumor model for in vitro drug penetration studies. Biomaterials 31:1180–1190PubMedCrossRefGoogle Scholar
  44. Paszek MJ, Zahir N, Johnson KR, Lakins JN, Rozenberg GI, Gefen A, Reinhart-King CA, Margulies SS, Dembo M, Boettiger D, Hammer DA, Weaver VM (2005) Tensional homeostasis and the malignant phenotype. Cancer Cell 8:241–254PubMedCrossRefGoogle Scholar
  45. Pinilla S, Alt E, Abdul Khalek FJ, Jotzu C, Muehlberg F, Beckmann C, Song YH (2009) Tissue resident stem cells produce CCL5 under the influence of cancer cells and thereby promote breast cancer cell invasion. Cancer Lett 284:80–85PubMedCrossRefGoogle Scholar
  46. Pollard JW (2004) Tumour-educated macrophages promote tumour progression and metastasis. Nat Rev Cancer 4:71–78PubMedCrossRefGoogle Scholar
  47. Ponzielli R, Katz S, Barsyte-Lovejoy D, Penn LZ (2005) Cancer therapeutics: targeting the dark side of Myc. Eur J Cancer 41:2485–2501PubMedCrossRefGoogle Scholar
  48. Raeber GP, Lutolf MP, Hubbell JA (2005) Molecularly engineered PEG hydrogels: a novel model system for proteolytically mediated cell migration. Biophys J 89:1374–1388PubMedCrossRefGoogle Scholar
  49. Richmond A, Su Y (2008) Mouse xenograft models vs GEM models for human cancer therapeutics. Dis Model Mech 1:78–82PubMedCrossRefGoogle Scholar
  50. Rizvanov AA, Yalvac ME, Shafigullina AK, Salafutdinov II, Blatt NL, Sahin F, Kiyasov AP, Palotas A (2010) Interaction and self-organization of human mesenchymal stem cells and neuro-blastoma SH-SY5Y cells under co-culture conditions: a novel system for modeling cancer cell micro-environment. Eur J Pharm Biopharm 76:253–259PubMedCrossRefGoogle Scholar
  51. Sahoo SK, Panda AK, Labhasetwar V (2005) Characterization of porous PLGA/PLA microparticles as a scaffold for three dimensional growth of breast cancer cells. Biomacromolecules 6:1132–1139PubMedCrossRefGoogle Scholar
  52. Serebriiskii I, Castello-Cros R, Lamb A, Golemis EA, Cukierman E (2008) Fibroblast-derived 3D matrix differentially regulates the growth and drug-responsiveness of human cancer cells. Matrix Biol 27:573–585PubMedCrossRefGoogle Scholar
  53. Shekhar MP, Werdell J, Santner SJ, Pauley RJ, Tait L (2001) Breast stroma plays a dominant regulatory role in breast epithelial growth and differentiation: implications for tumor development and progression. Cancer Res 61:1320–1326PubMedGoogle Scholar
  54. Sutherland RM (1988) Cell and environment interactions in tumor microregions: the multicell spheroid model. Science 240:177–184PubMedCrossRefGoogle Scholar
  55. Talukdar S, Mandal M, Hutmacher DW, Russell PJ, Soekmadji C, Kundu SC (2011) Engineered silk fibroin protein 3D matrices for in vitro tumor model. Biomaterials 32:2149–2159PubMedCrossRefGoogle Scholar
  56. Tian B, Li Y, Ji XN, Chen J, Xue Q, Ye SL, Liu YK, Tang ZY (2005) Basement membrane proteins play an active role in the invasive process of human hepatocellular carcinoma cells with high metastasis potential. J Cancer Res Clin Oncol 131:80–86PubMedCrossRefGoogle Scholar
  57. Verbridge SS, Choi NW, Zheng Y, Brooks DJ, Stroock AD, Fischbach C (2010) Oxygen-controlled three-dimensional cultures to analyze tumor angiogenesis. Tissue Eng A 16:2133–2141CrossRefGoogle Scholar
  58. Villanueva I, Weigel CA, Bryant SJ (2009) Cell-matrix interactions and dynamic mechanical loading influence chondrocyte gene expression and bioactivity in PEG-RGD hydrogels. Acta Biomater 5:2832–2846PubMedCrossRefGoogle Scholar
  59. Webber MM, Bello D, Kleinman HK, Hoffman MP (1997) Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. Carcinogenesis 18:1225–1231PubMedCrossRefGoogle Scholar
  60. World Health Organization. Cancer. http://www.who.int/cancer/en. Accessed 08 March 2011

Copyright information

© The International CCN Society 2011

Authors and Affiliations

  • Agata Nyga
    • 1
    • 2
  • Umber Cheema
    • 2
    • 3
  • Marilena Loizidou
    • 1
    • 2
    • 4
  1. 1.Centre for Nanotechnology, Biomaterials and Tissue EngineeringUniversity College LondonLondonUK
  2. 2.UCL Division of Surgery & Interventional ScienceUniversity College LondonLondonUK
  3. 3.Tissue Repair and Engineering Centre, Institute of Orthopaedics and Musculoskeletal ScienceUniversity College LondonLondonUK
  4. 4.UCL Division of Surgery and Interventional ScienceRoyal Free Hospital, 9th floorLondonUK

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