Abstract
In vitro assays with endothelial cells (EC) cultured on three-dimensional gel recapitulate several aspects of vascular morphogenesis and pathological angiogenesis. The two most used in vitro assays of vascular morphogenesis are the tube formation on extracellular matrix gel and the sprouting from EC spheroids. Tube formation assay measures the ability of EC, plated on gel derived from reconstituted basement membrane, to form capillary-like structures. Sprouting assay is based on spheroids of EC, embedded in collagen gel and stimulated with angiogenic factors, which originate a complex network of capillary-like structures invading the gel.
Both these assays can be exploited for antiangiogenic drug screening and gene function analysis during vascular morphogenesis.
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References
Folkman J (1976) The vascularization of tumors. Sci Am 234:58–64
Risau W (1997) Mechanisms of angiogenesis. Nature 386:671–674
Carmeliet P (2000) Mechanisms of angiogenesis and arteriogenesis. Nat Med 6:389–395
Folkman J, Haudenschild C (1980) Angiogenesis in vitro. Nature 288:551–556
Jain RK, Schlenger K, Hockel M, Yuan F (1997) Quantitative angiogenesis assays: progress and problems. Nat Med 3:1203–1208
Auerbach R, Auerbach W, Polakowski I (1991) Assays for angiogenesis: a review. Pharmacol Ther 51:1–11
Conway EM, Collen D, Carmeliet P (2001) Molecular mechanisms of blood vessel growth. Cardiovasc Res 49:507–521
Hanahan D (1997) Signaling vascular morphogenesis and maintenance. Science 277:48–50
Ausprunk DH, Knighton DR, Folkman J (1975) Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Role of host and preexisting graft blood vessels. Am J Pathol 79:597–618
Korff T, Krauss T, Augustin HG (2004) Three-dimensional spheroidal culture of cytotrophoblast cells mimics the phenotype and differentiation of cytotrophoblasts from normal and preeclamptic pregnancies. Exp Cell Res 297:415–423
Maheshwari RK, Srikantan V, Bhartiya D, Kleinman HK, Grant DS (1991) Differential effects of interferon gamma and alpha on in vitro model of angiogenesis. J Cell Physiol 146:164–169
Madri JA, Pratt BM (1986) Endothelial cell-matrix interactions: in vitro models of angiogenesis. J Histochem Cytochem 34:85–91
Primo L, Seano G, Roca C, Maione F, Gagliardi PA, Sessa R et al (2010) Increased expression of alpha6 integrin in endothelial cells unveils a proangiogenic role for basement membrane. Cancer Res 70:5759–5769
Korff T, Augustin HG (1998) Integration of endothelial cells in luticellualr spheroids prevent apoptosis and induces differentiation. J Cell Biol 143:1341–1352
Sessa R, Seano G, di Blasio L, Gagliardi PA, Isella C, Medico E et al (2012) The miR-126 regulates angiopoietin-1 signaling and vessel maturation by targeting p85beta. Biochim Biophys Acta 1823:1925–1935
Bussolino F, Di Renzo MF, Ziche M, Bocchietto E, Olivero M, Naldini L et al (1992) Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth. J Cell Biol 119:629–641
Serini G, Ambrosi D, Giraudo E, Gamba A, Preziosi L, Bussolino F (2003) Modeling the early stages of vascular network assembly. EMBO J 22:1771–1779
Seano G, Chiaverina G, Gagliardi PA, di Blasio L, Sessa R, Bussolino F et al (2013) Modeling human tumor angiogenesis in a three-dimensional culture system. Blood 121:e129–e137
Acknowledgment
“Associazione Italiana per la Ricerca sul Cancro” (AIRC: grants 14284, 14635, 10133, 12182); the University of Torino-Compagnia di San Paolo: RETHE, grant-ORTO11RKTW (to F.B.); MIUR Project Newton (grant FIRB RBAP11BYNP); Telethon, Italy (grant GEP13015); FP7 “BILOBA” Grant (contract 318035)
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di Blasio, L., Bussolino, F., Primo, L. (2015). Three-Dimensional In Vitro Assay of Endothelial Cell Invasion and Capillary Tube Morphogenesis. In: Ribatti, D. (eds) Vascular Morphogenesis. Methods in Molecular Biology, vol 1214. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1462-3_4
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DOI: https://doi.org/10.1007/978-1-4939-1462-3_4
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