Publication Database

Vimentin knock-out and its impact on angiogenesis in vitro (2019)

Art

Poster

Autoren

Herre, Christina (WE 1)

Nshdejan, Arpenik (WE 1)

Bahramsoltani, Mahtab (WE 1)

Kongress

12. Doktorandensymposium & DRS Präsentationsseminar "Biomedical Sciences"

Berlin, 27.09.2019

Quelle

Doktorandensymposium & DRS Präsentationsseminar "Biomedical Sciences"

Berlin: Mensch und Buch Verlag, 2019

ISBN: 978-3-96729-006-6

Sprache

Englisch

Kontakt

Institut für Veterinär-Anatomie

Koserstr. 20
14195 Berlin
+49 30 838 75784
anatomie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

Introduction:
Angiogenesis is defined as the growth and remodeling of new blood vessels due to proliferation and differentiation of endothelial cells (ECs) of pre-existing capillaries. It plays a significant role in ischemic diseases, tumor growth and metastasis. Therefore, many in vivo and in vitro angiogenic assays have been developed. To reduce time, cost and the number of animal experiments, recent research has focused on optimizing in vitro assays. Hence, an in vitro model of angiogenesis was established providing a method to quantify angiogenesis by dividing the process into six defined stages (Bahramsoltani et al. Clin Hemorheol Microcirc 2010; 55, 255-269). ECs display different angiogenic potencies, categorized into angiogenic and non-angiogenic. However, reliable markers for the characterization of the different ECs are non-existent. During previous studies, eight specific proteins were identified as potential markers (Bahramsoltani et al. Clin Hemorheol Microcirc 2013; 55, 255-269). This study focuses on the examination of the angiogenic potency of one of these proteins, Vimentin.

Material and Methods:
A long-term cultivation of human microvascular endothelial cells derived from foreskin (HDMEC) was performed. Angiogenesis stimulation was induced through a proangiogenic medium. Subcultures were infected with specifically designed short hairpin RNAs (shRNAs), which induced the knock-out of the expression of Vimentin. Morphological changes were captured and assigned to the defined stages throughout cultivation. Furthermore, the incorporation of the shRNA plasmid was detected due to the biosensor green fluorescent protein.

Results:
During the investigation period, non-transfected ECs ran through the six defined stages of angiogenesis, chronologically. In contrast, infected cells showed a higher rate of cell death. While sprouting was observed, cells were unable to proceed to further angiogenic stages.

Conclusion:
Vimentin plays a major role in angiogenesis. Gene silencing inhibits cells from completing all stages of angiogenesis. Current efforts focus on gene silencing of a second protein found, Myozenin-2.