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Poster display

3837 - A multicellular 3D cell culture model for investigation of endothelial cell migration


10 Oct 2016


Poster display


Arno Amann


Annals of Oncology (2016) 27 (6): 526-544. 10.1093/annonc/mdw392


A. Amann1, M. Zwierzina2, G. Gamerith3, S. Koeck3, E. Lorenz4, H. Zwierzina3, J. Kern4

Author affiliations

  • 1 Innere Medizin, Medizinische Universität Innsbruck, 6020 - Innsbruck/AT
  • 2 Department Of Plastic, Reconstructive And Aesthetic Surgery, Austria, Medizinische Universität Innsbruck, 6020 - Innsbruck/AT
  • 3 Haematology And Oncology, Medizinische Universität Innsbruck, 6020 - Innsbruck/AT
  • 4 Oncotyrol - Center For Personalized Cancer Medicine, Oncotyrol - Center for Personalized Cancer Medicine, 6020 - Innsbruck/AT


Abstract 3837


We are experiencing a transition from disease to target-oriented therapy due to the increasing understanding of the mechanisms relevant to the genesis of cancer. One major hurdle for the development of these targeted therapeutic regimens, however, is the limited availability of predictive in vitro models. We present data that highlights the differences of RNA expression of in vivo like 3D microtissues consisting of tumour cells, fibroblasts and two different endothelial cell lines compared to normal 2D cell culture conditions.


96-well hanging drop microtiter plates (InSphero AG, Zürich, Switzerland) were applied for the production of 3D mono-, co- and tri-cultures including the human lung cancer cell lines A549 or Colo699 alone or in combination with a human lung fibroblast cell line (SV-80) and either a human umbilical vein endothelial cell line (HUVEC) or the primary human lung microvascular endothelial cell line (HMVEC-L). Tumour endothelial spheroid aggregation was displayed immunohistochemically (IHC) by protein expression of e-cadherin, CD31, von Willebrand factor (vWF) and α-muscle actin (α-SMA). RNA expression profiling by Affymetrix chip analysis was performed for multicellular 3D microtissues and 2D cultured cell lines. Bevacizumab was added in different doses and inhibition of endothelial cell migration and drug related toxicity was displayed either by flow cytometry or IHC.


In microtissues, endothelial cells aggregated in coherent tube-like structures preferentially in the fibroblast consisting core of all microtissues. However, inhibition of vascular endothelial growth (VEGF) factor by bevacizumab led to an in part blockade of endothelial cell migration into the microtissues. Nevertheless, no toxic effect of this drug was displayed either on tumour cells, fibroblasts or endothelial cells. RNA expression profiles revealed a high number of regulated genes in tri-cultures when compared to microtissues only consisting of mono- or co-cultures or to traditional 2D cultivated cells.


In this work, we demonstrate a functional multicellular model consisting of tumour cells, fibroblasts and endothelial cells that allows the investigation of anti-angiogenic drugs.

Clinical trial identification

Legal entity responsible for the study

Medical University Innsbruck Tirol Kliniken


Medical University Innsbruck Tirol Kliniken


All authors have declared no conflicts of interest.

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