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World Journal of Surgery

, Volume 31, Issue 4, pp 654–663 | Cite as

In Vitro Models of Angiogenesis

  • Areck A. Ucuzian
  • Howard P. Greisler
Article

Abstract

Neovascularization can be categorized into two general processes: vasculogenesis and angiogenesis. Angiogenesis is the formation of new capillaries from pre-existing vessels, requiring growth factor driven recruitment, migration, proliferation, and differentiation of endothelial cells (ECs). Complex cell–cell and cell-extracellular matrix (ECM) interactions contribute to this process, leading finally to a network of tube-like formations of endothelial cells supported by surrounding mural cells. The study of angiogenesis has broad clinical implications in the fields of peripheral and coronary vascular disease, oncology, hematology, wound healing, dermatology, and ophthalmology, among others. As such, novel, clinically relevant models of angiogenesis in vitro are crucial to the understanding of angiogenic processes. We highlight some of the advances made in the development of these models, and discuss the importance of incorporating the three-dimensional cell-matrix and EC–mural cell interactions into these in vitro assays of angiogenesis. This review also discusses our own 3-D angiogenesis assay and some of the in vitro results from our lab as they relate to therapeutic neovascularization and tissue engineering of vascular grafts.

Keywords

Vascular Endothelial Growth Factor Fibroblast Growth Factor Mural Cell Angiogenesis Model Growth Factor Delivery 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by grants (to H.P.G.) from the National Institutes of Health (NIH R01-HL41272) and the Department of Veteran’s Affairs.

References

  1. 1.
    Kurz H, Burri PH, Djonov VG. Angiogenesis and vascular remodeling by intussusception: from form to function. News Physiol Sci 2003;18:65–70PubMedGoogle Scholar
  2. 2.
    Scholz D, Cai WJ, Schaper W. Arteriogenesis, a new concept of vascular adaptation in occlusive disease. Angiogenesis 2001;4:247–257PubMedCrossRefGoogle Scholar
  3. 3.
    Risau W. Mechanism of angiogenesis. Nature 1997;386:671–674PubMedCrossRefGoogle Scholar
  4. 4.
    Murray CJ, Lopez AD. Mortality by cause for eight regions of the world: Global Burden of Disease Study. Lancet 1997;349:1269–1276PubMedCrossRefGoogle Scholar
  5. 5.
    American Heart Association. http://www.americanheart.org 11/10/05
  6. 6.
    World Health Organization. http://www.who.int. 11/10/05
  7. 7.
    Baffour R, Berman J, Garb JL, et al. Enhanced angiogenesis and growth of collaterals by in vivo administration of recombinant basic fibroblast growth factor in a rabbit model of acute lower limb ischemia. J Vasc Surg 1992;16:181–191PubMedCrossRefGoogle Scholar
  8. 8.
    Pu LQ, Sniderman AD, Brassard R, et al. Enhanced revascularization of the ischemic limb by angiogenic therapy. Circulation 1993;88:208–215PubMedGoogle Scholar
  9. 9.
    Holmgran L, O’Reilly MS, Folkman J. Dormancy of micrometastases: balanced proliferation and apoptosis in the presence of angiogenesis suppression. Nat Med 1995;1:149–153CrossRefGoogle Scholar
  10. 10.
    Fernandez A, Udagawa T, Schwesinger C, et al. Angiogenic potential of prostate carcinoma cells overexpressing Bcl-2. J Natl Cancer Inst 2001;93:208–213PubMedCrossRefGoogle Scholar
  11. 11.
    Folkman J. Angiogenesis. Annu Rev Med 2006;57:1–18PubMedCrossRefGoogle Scholar
  12. 12.
    Folkman J, Haudenschild C. Angiogenesis in vitro. Nature 1980;288:551–556PubMedCrossRefGoogle Scholar
  13. 13.
    Auerbach R, Akhtar N, Lewis RL, et al. Angiogenesis assays: problems and pitfalls. Cancer Metastasis Rev 2000;19:167–172PubMedCrossRefGoogle Scholar
  14. 14.
    Auerbach R, Lewis R, Shinners B, et al. Angiogenesis assays: a critical overview. Clin Chem 2003;49:32–40PubMedCrossRefGoogle Scholar
  15. 15.
    Jain RK, Schlenger K, Hockel M, et al. Quantitative angiogenesis assays: progress and problems. Nat Med 1997;3:1203–1208PubMedCrossRefGoogle Scholar
  16. 16.
    Pepper MS. Manipulating angiogenesis. From basic science to the bedside. Arterioscler Thromb Vasc Biol 1997;17:605–619PubMedGoogle Scholar
  17. 17.
    Lawley TJ, Kubota Y. Induction to morphologic differentiation of endothelial cells in culture. J Invest Dermatol 1989;93:59S–61SPubMedCrossRefGoogle Scholar
  18. 18.
    Kanzawa S, Endo H, Shioya N, et al. Improved in vitro angiogenesis model by collagen density reduction and the use of type III collagen. Ann Plast Surg 1993;30:244–251PubMedCrossRefGoogle Scholar
  19. 19.
    Davis GE, Bayless KJ, Mavila A. Molecular basis of endothelial cell morphogenesis in three-dimensional extracellular matrices. Anat Rec 2002;268:252–275PubMedCrossRefGoogle Scholar
  20. 20.
    Bishop ET, Bell GT, Bloor S, et al. An in vitro model of angiogenesis: basic features. Angiogenesis 1999;3:335–344PubMedCrossRefGoogle Scholar
  21. 21.
    Donovan D, Brown NJ, Bishop ET, et al. Comparison of three in vitro human “angiogenesis” assays with capillaries formed in vivo. Angiogenesis 2001;4:113–121PubMedCrossRefGoogle Scholar
  22. 22.
    Griffith LG, Swartz MA. Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 2006;7:211–224PubMedCrossRefGoogle Scholar
  23. 23.
    Roskelley CD, Bissell MJ. Dynamic reciprocity revisited: a continuous, bi-directional flow of information between cells and the extracellular matrix regulates mammary epithelial cell function. Biochem Cell Biol 1995;73:391–397PubMedCrossRefGoogle Scholar
  24. 24.
    Ross R. Molecular and mechanical synergy: cross-talk between integrins and growth factor receptors. Cardiovasc Res 2004;63:381–390PubMedCrossRefGoogle Scholar
  25. 25.
    Nicosia RF, Otinetti A. Growth of microvessels in serum-free matrix culture of rat aortas. A quantitative assay of angiogenesis in vitro. Lab Invest 1990;63:115–122PubMedGoogle Scholar
  26. 26.
    Nicosia RF, Lin YJ, Hazelton D, et al. Endogenous regulation of angiogenesis in the rat aorta modal. Role of vascular endothelial growth factor. Am J Pathol 1997;151:1379–1386PubMedGoogle Scholar
  27. 27.
    Muthukkaruppan VR, Shinners BL, Lewis R, et al. The chick embryo aortic arch assay: a new, rapid, quantifiable in vitro method for testing the efficacy of angiogenic and anti-angiogenic factors in a three-dimensional, serum-free organ culture system. Proc Am Assoc Cancer Res 2000;41:65Google Scholar
  28. 28.
    Brown KJ, Maynes SF, Bezos A, et al. A novel in vitro angiogenesis assay. Lab Invest 1996;75: 539–555PubMedGoogle Scholar
  29. 29.
    Callow AD. Current status of vascular grafts. Surg Clin North Am 1983;62:501–513Google Scholar
  30. 30.
    Clagget GP, Colman RW, Marder VJ, et al., eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice, Philadelphia, JB Lippincott Co, 1987;1348–1365Google Scholar
  31. 31.
    Crawford ES, Bomberger RA, Glaeser DH, et al. Aortoiliac occlusive disease: factors influencing survival and function following reconstructive operation over a twenty-five year period. Surgery 1981;90:1055–1066PubMedGoogle Scholar
  32. 32.
    Versaci F, Gaspardone A, Tomai F, et al. A comparison of coronary-artery stenting with angioplasty for isolated stenosis of the proximal left anterior descending coronary artery. N Engl J Med 1997;336:817–822PubMedCrossRefGoogle Scholar
  33. 33.
    Rodriguez AE, Santaera O, Iarribau M, et al. Coronary stenting decreases restenosis in lesions with early loss in luminal diameter 24 hours after successful PTCA. Circulation 1995;91:1397–1402PubMedGoogle Scholar
  34. 34.
    Fischman DL, Leon MB, Baim DS, et al. A randomized comparison of coronary-stent placement and balloon angioplasty in the treatment of coronary artery disease. N Engl J Med 1994;331:496–501PubMedCrossRefGoogle Scholar
  35. 35.
    Zarge JI, Huang P, Husak V, et al. Fibrin glue containing fibroblast growth factor type 1 and heparin with autologous endothelial cells reduces intimal hyperplasia in a canine carotid artery balloon injury model. J Vasc Surg 1997;25:840–848PubMedCrossRefGoogle Scholar
  36. 36.
    Xue L, Shireman P, Hampton B, et al. The cysteine-free fibroblast growth factor-1 mutant induces heparin-independent proliferation of endothelial cells and smooth muscle cells. J Surg Res 2000;92:255–260PubMedCrossRefGoogle Scholar
  37. 37.
    Shireman PK, Xue L, Maddox E, et al. The S 130K fibroblast growth factor-1 mutant induces heparin-independent proliferation and is resistant to thrombin degradation in fibrin glue. J Vasc Surg 2000;31:382–390PubMedCrossRefGoogle Scholar
  38. 38.
    Erzurum VZ, Bian JF, Husak VA, et al. R136K fibroblast growth factor-1 mutant induces heparin-independent migration of endothelial cells through fibrin glue. J Vasc Surg 2003;37:1075–1081PubMedCrossRefGoogle Scholar
  39. 39.
    Vernon RB, Sage EH. A novel, quantitative model for study of endothelial cell migration and sprout formation within 3-dimensional collagen matrices. Microvasc Res 1999;57:118–133PubMedCrossRefGoogle Scholar
  40. 40.
    Pepper MS, Ferrara N, Orci L, et al. Potent synergism between vascular endothelial growth factor and basic fibroblast growth factor in the induction of angiogenesis in vitro. Biochem Biophys Res Commun 1992;189:824–831PubMedCrossRefGoogle Scholar
  41. 41.
    Xue L, Greisler HP. Angiogenic effect of fibroblast growth factor-1 and vascular endothelial growth factor and their synergism in a novel in vitro quantitative fibrin-based 3-dimensional angiogenesis system. Surgery 2002;132:259–267PubMedCrossRefGoogle Scholar
  42. 42.
    Gospodarowicz D, Cheng J. Heparin protects basic and acidic FGF from inactivation. J Cell Physiol 1986;128:475–484PubMedCrossRefGoogle Scholar
  43. 43.
    Sahni A, Sporn LA, Francis CW. Potentiation of endothelial cell proliferation by fibrin(ogen)-bound fibroblast growth factor-2. J Biol Chem 1999;274:14936–14941PubMedCrossRefGoogle Scholar
  44. 44.
    Chalupowicz DG, Chowdhury ZA, Bach TL, et al. Fibrin II induces endothelial cell capillary tube formation. J Cell Biol 1995;130:207–215PubMedCrossRefGoogle Scholar
  45. 45.
    Shireman PK, Hampton B, Burgess WH, et al. Modulation of vascular cell growth kinetics by local cytokine delivery from fibrin glue suspensions. J Vasc Surg 1999;29:852–861PubMedCrossRefGoogle Scholar
  46. 46.
    Shireman PK, Greisler HP. Mitogenicity and release of vascular endothelial growth factor with and without heparin from fibrin glue. J Vasc Surg 2000;31:936–943PubMedCrossRefGoogle Scholar
  47. 47.
    Gu F, Amsden B, Neufeld R. Sustained delivery of vascular endothelial growth factor with alginate beads. J Control Release 2004;96:463–472PubMedCrossRefGoogle Scholar
  48. 48.
    Brey EM, Uriel S, Greisler HP, et al. Therapeutic neovascularization: contribution from bioengineering. Tissue Eng 2005;11:567–584PubMedCrossRefGoogle Scholar
  49. 49.
    Epstein SE, Kornowski R, Fuchs S, et al. Angiogenesis therapy: amidst the hype, the neglected potential for serious side effects. Circulation 2001;104:115–119PubMedGoogle Scholar
  50. 50.
    Uriel S, Brey EM, Greisler HP, et al. Sustained low levels of FGF-1 promote persistent microvascular network formation. Am J Surg 192:604–609Google Scholar
  51. 51.
    Ozawa CR, Banfi A, Glazer NL, et al. Microenvironmental VEGF concentration, not total dose, determines a threshold between normal and aberrant angiogenesis. J Clin Invest 2004;113:516–527PubMedCrossRefGoogle Scholar
  52. 52.
    Hirschi KK, Rohovsky SA, D’Amore PA, et al. PDGF, TGF-beta, and heterotypic cell–cell interactions mediate endothelial cell-induced recruitment of 10T1/2 cells and their differentiation to a smooth muscle fate. J Cell Biol 1998;141:805–814PubMedCrossRefGoogle Scholar
  53. 53.
    Hirschi KK, Rohovsky SA, Beck LH, et al. Endothelial cells modulate the proliferation of mural cell precursors via platelet-derived growth factor-BB and heterotypic cell contact. Circ Res 1999;84:298–305PubMedGoogle Scholar
  54. 54.
    Nackman GB, Fillinger MF, Shafritz R, et al. Flow modulates endothelial regulation of smooth muscle cell proliferation: a new model. Surgery 1998;124:353–360PubMedGoogle Scholar
  55. 55.
    Heydarkhan-Hagvall S, Helenius G, Johansson BR, et al. Co-culture of endothelial cells and smooth muscle cells affects gene expression of angiogenic factors. J Cell Biochem 2003;89:1250–1259PubMedCrossRefGoogle Scholar
  56. 56.
    Darland DC, D’Amore PA. TGF beta is required for the formation of capillary-like structures in three-dimensional co-cultures of 10T1/2 and endothelial cells. Angiogenesis 2001;4:11–20PubMedCrossRefGoogle Scholar
  57. 57.
    Korff T, Kimmina S, Martiny-Baron G, et al. Blood vessel maturation in a 3-dimensional spheroidal co-culture model: direct contact with smooth muscle cells regulates endothelial cell quiescence and abrogates VEGF responsiveness. FASEB J 2001;15:447–5757PubMedCrossRefGoogle Scholar
  58. 58.
    Korff T, Augustin HG. Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. J Cell Sci 1999;112:3249–3258PubMedGoogle Scholar
  59. 59.
    Niklason LE, Gao J, Abbott WM, et al. Functional arteries grown in vitro. Science 1999;284:489–493PubMedCrossRefGoogle Scholar
  60. 60.
    Kaushal S, Amiel GE, Guleserian KJ, et al. Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo. Nat Med 2001;7:1035–1040PubMedCrossRefGoogle Scholar

Copyright information

© Société Internationale de Chirurgie 2007

Authors and Affiliations

  1. 1.Department of SurgeryLoyola University Medical CenterMaywoodUSA
  2. 2.Department of Cell Biology, Neurobiology, and AnatomyLoyola University Medical CenterMaywoodUSA
  3. 3.Department of Surgery and Research ServiceHines V.A. HospitalHinesUSA

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