DEVELOPING AN IN VITRO THREE DIMENSIONAL DYNAMIC BLOOD BRAIN BARRIER MODEL AND INVESTIGATING THE MODEL’S PERMEABILITY
Project no. 1233 · Status: Kapanmış · 2016
Summary
Today there are several BBB models frequently used in drug screening, targeting and development studies, but none of those are able to properly imitate the permeability of this special barrier. Due to ethical limitations to the use of experimental animals and the fact that the barrier system in animals is different than that in humans make the development of a realistic model that can be safely and reliably used in drug targeting studies and investigating factors can affect BBB essential. The aim of this thesis is the construction of a reliable in vitro BBB model mimicking physiological BBB in terms of cellular organization and mechanobiology. With this aim, as a replacement of the basal membrane, bacterial cellulose (BC), which is previously shown to imitate extracellular matrix was used for developing the new three-dimensional (3D) BBB system. Brain microvascular endothelial cells (BMECs) were seeded in luminal section and astrocytes and brain microvascular pericytes (BMPCs) were seeded in abluminal section of the membrane. Co-cultivation of these three cell types is essential in order for the BBB model to be more realistic by reducing permeability. The cells were cultivated both on a two-dimensional (2D) static BBB model and in the 3D BBB reactor system, which is designed and constructed in order to simulate the physiological blood flow, shear stress and circumferential stress caused by the heart beat on brain capillaries; and amount of expression of tight and adherence junction proteins were determined, morphological changes and vitality were examined, and permeability of the model was examined in molecular level and determined by measuring trans-endothelial electrical resistance (TEER).