A dynamic culture platform enhances the efficiency of the 3D HUVEC‐based tube formation assay

• Published in: Biotechnology and bioengineering Vol. 117; no. 3; pp. 789 - 797
• Main Authors: Lovecchio, Joseph, Pannella, Micaela, Giardino, Luciana, Calzà, Laura, Giordano, Emanuele
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-03-2020
• Subjects: Angiogenesis; Bathing; Biological models (mathematics); Bioreactor system; Bioreactors; Biotechnology; Cell culture; Cell Culture Techniques - methods; Cells, Cultured; Diffusion rate; drug discovery; Edetic acid; Endothelial cells; Ethylenediaminetetraacetic acids; Finite Element Analysis; finite element modeling; Human Umbilical Vein Endothelial Cells - cytology; Humans; HUVECs; Neovascularization, Physiologic - physiology; Perfusion; periodic perfusion; Physiology; Tissues; tube formation assay; Umbilical vein; Bioreactor system; finite element modeling; drug discovery; tube formation assay; HUVECs; periodic perfusion
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.27227

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.
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 matrix.