Publikationsdatenbank

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

Bandzählung: 386

Heftzählung: 2

Seiten: 167 – 75

ISSN: 1431-6730

Sprache

Englisch

Verweise

Pubmed: 15843161

Kontakt

Institut für Veterinär-Anatomie

Koserstr. 20
14195 Berlin
+49 30 838 75784
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.