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    Morphology and transfection study of human microvascular endothelial cell angiogenesis: an in vitro three-dimensional model (2005)

    Art
    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Autoren
    Lienau, Jasmin
    Kaletta, Cortina
    Teifel, Michael
    Naujoks, Kurt
    Bhoola, Kanti
    Plendl, Johanna (WE 1)
    Quelle
    Biological chemistry; 386(2) — S. 167–75
    ISSN: 1431-6730
    Sprache
    Englisch
    Verweise
    Pubmed: 15843161
    Kontakt
    Institut für Veterinär-Anatomie

    Koserstr. 20
    14195 Berlin
    Tel.+49 30 838 53555 Fax.+49 30 838-53480
    email:anatomie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    Microvascular endothelial cells from human neonatal foreskin were grown in vitro until a three-dimensional network of capillary-like structures was formed. All stages of the angiogenic cascade could be observed in this in vitro model, including the formation of an internal lumen. The microscopy focused on morphology, formation of an internal lumen, role of the extracellular matrix, polarity of the cells, and the time-course of the angiogenic cascade. Bright-field microscopy revealed cells arranged circularly side by side and the internal lumen of capillary-like structures was verified by electron microscopy. Immunolabeling revealed a peritubular localization of collagen IV. Reporter gene expression after the formation of capillary-like structures was marginally higher than control expression, but clearly lower than the expression of cells at the stage of proliferation. Highest transfection efficiencies were obtained using vectors with the CMV promoter and the long fragment of the Ets-1 promoter. This is a first study of transfection efficiencies mapped for stages of in vitro angiogenesis. We describe here the morphological features of a long-term in vitro model of angiogenesis of human microvascular endothelial cells that could be used for transfection studies, without the provision of an extracellular matrix substrate. The cells self-create their own extracellular matrix to proliferate and form a three-dimensional network of capillary-like structures with an internal lumen.