Improved in vitro angiogenesis model by collagen density reduction and the use of type III collagen.
Angiogenesis was examined by a three-dimensional model in vitro, using human umbilical vein endothelial cells (HUVECs) cultured in a collagen gel. An abundant capillary-like network with a lumen structure was identified histologically and shown to have formed at a collagen density of 0.05% or 0.10% instead of 0.15%, for either type I or type III collagen. At the same density, type III collagen induced a capillary-like network with HUVECs at an earlier stage of culture than type I collagen. In a two-dimensional culture, HUVECs were viable and proliferated to a great extent on the culture dish coated with collagen. This was particularly so in the case of type III collagen at lower density (5 ng/cm2). Type III collagen at a sufficiently low density is thus shown useful for studying angiogenesis in vitro. The capillary-like network that formed in the three-dimensional culture appeared somewhat labile, but became stable with the continuous addition of endothelial cell growth supplement and increase in the plating number of HUVECs.
Kanzawa
S
S
Department of Plastic Surgery, Kitasato University School of Medicine, Kanagawa, Japan.
Endo
H
H
Shioya
N
N
D002196:Capillaries / Q000473:pathology; D002454:Cell Differentiation / Q000502:physiology; D003094:Collagen / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000473:pathology*; D005109:Extracellular Matrix / Q000473:pathology; D006801:Humans; D066298:In Vitro Techniques; D009389:Neovascularization, Pathologic / Q000473:pathology*; D014471:Umbilical Veins / Q000473:pathology
D003094:Collagen