Establishing hiPSC-derived brain microvascular endothelial cells (BMVEC) for in-vitro modelling of bubble-assisted focused ultrasound (BAFUS) disruption of the blood brain barrier (BBB)
The escalating incidence of neurological disorders including cancers due to increased life expectancy necessitates the development of more effective neurotherapeutics. A significant hurdle in this pursuit is the human blood-brain barrier (hBBB), which impedes the delivery of almost 100% of large-molecule drugs and 98% of small-molecule drugs from blood to the brain. Despite various attempts to overcome this issue in the last two decades, the challenge still remains. Recently, bubble-assisted focused ultrasound (BAFUS), has proven to be able to temporarily open the BBB for drug delivery at targeted brain locations and is currently in clinical trials. An in-vitro model of the technique should improve the drug delivery pipeline and speed up the optimization of personalized drug delivery and dosing. Previously, our lab has developed a ‘US-transparent BBB organ-on-chip model’, and preliminary BBB disruption and recovery have been observed. However, the cellular barrier was a GI epithelial Caco-2 cell barrier with a Trans-Epithelial/Endothelial Electric Resistance (TEER) of 300-400 Ω-cm2, which is not in-vivo BBB specific.
In this project, we aim to use human induced Pluripotent Stem Cell (hiPSC)-derived Brain Microvascular Endothelial Cells (BMVECs) to reproduce the BBB for highly predictive and patient-specific in-vitro modeling. After reagent selection, the iPS(IMR90)-4 line was successfully cultured and banked. Morphology has been used to characterize iPSCs and differentiated cells. Cell adhesion difference has been observed for different culturewares. ReLeSR was tested for differential dissociation of iPSCs. The BMEC differentiation is currently in process with an expected barrier TEER > 2000 Ω-cm2. If successful, we expect the iPSC-derived BMEC BBB to be a critical enabler for establishing predictive and personalized BAFUS BBB disruption in-vitro models.