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A dynamic culture platform enhances the efficiency of the 3D HUVEC-based tube formation assay |
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| Authors: | Joseph Lovecchio Micaela Pannella Luciana Giardino Laura Calzà Emanuele Giordano |
| Institution: | 1. Health Sciences and Technologies—Interdepartmental Center for Industrial Research, University of Bologna, Bologna, Italy;2. Health Sciences and Technologies—Interdepartmental Center for Industrial Research, University of Bologna, Bologna, Italy
Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy;3. Health Sciences and Technologies—Interdepartmental Center for Industrial Research, University of Bologna, Bologna, Italy Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy;4. Health Sciences and Technologies—Interdepartmental Center for Industrial Research, University of Bologna, Bologna, Italy Laboratory of Cellular and Molecular Engineering “Silvio Cavalcanti”—Department of Electrical, Electronic and Information Engineering “Guglielmo Marconi” (DEI), University of Bologna, Cesena, Italy Advanced Research Center on Electronic Systems “Ercole De Castro”, University of Bologna, Bologna, Italy |
| Abstract: | Cell-based in vitro biological models traditionally use monolayer cell cultures grown over plastic surfaces bathing in static media. Higher fidelity to a natural biological tissue is expected to result from growing the cells in a three-dimensional (3D) matrix. However, due to the decreased rate of diffusion inherent to increased distances within a tridimensional space, proper fluidic conditions are needed in this setting to better approximate a physiological environment. To this aim, we here propose a prototypal dynamic cell culture platform for the automatic medium replacement, via periodic perfusion flow, in a human umbilical vein endothelial cell (HUVECs) culture seeded in a Geltrex™ matrix. A state-of-the-art angiogenesis assay performed in these dynamic conditions showed sizable effects with respect to conventional static control cultures, with significantly enhanced pro-(dual antiplatelet therapy DAPT]) and anti-(EDTA) angiogenic compound activity. In particular, dynamic culture conditions (a) enhance the 3D-organization of HUVECs into microtubule structure; (b) accelerate and improve endothelial tube formation by HUVECs in the presence of DAPT; (c) are able to completely revert the blocking effects of EDTA. These evidence emphasize the need of setting proper fluidic conditions for a better approximation of a physiological environment as an appropriate evolution of current cell culture paradigms. |
| Keywords: | Bioreactor system drug discovery finite element modeling HUVECs periodic perfusion tube formation assay |