Real-time patch-clamp sensing of transport across a barrier tissue
| DWPI Title: Measuring electrical signal from cultured cell layer, for quantifying vesicle fusion to membranes through measurement of capacitance, for testing ability of agent to cross barrier layer, comprises culturing barrier cells, adding agent and measuring electrical signals where barrier cell are hPSCs |
| Abstract: The present disclosure relates to methods, apparatuses, and systems to measure one or more electrical signals from a cultured cell layer. In some embodiments, such electrical signals can be indicative of transport or lack of transport of an agent through the cell layer. |
| Use: Method for measuring electrical signals from a cultured cell layer, for quantifying vesicle fusion to membranes through measurement of capacitance, with a resolution sufficient for detecting individual vesicles, for testing ability of agent to cross the barrier layer. |
| Advantage: The method allows spatial and temporal resolution of electrical signals being generated by the barrier layer. |
| Novelty: Measuring electrical signals from a cultured cell layer comprises (a) culturing barrier cells on a first surface of a substrate and within a first chamber, where the substrate comprises pores disposed within the substrate, and pore is a micropore or a nanopore and forming a seal between pore and barrier cell; (b) adding an agent to the first chamber; and (c) measuring electrical signals in first chamber or second chamber in fluidic communication with one pore, where the electrical signals is indicative of transport or lack of transport of the agent through the barrier cell, the barrier cells comprises a layer of cells where the barrier cell remains functionally active and intact during the measuring step and the barrier cells comprises a co-culture of a barrier cell selected from group consisting of brain microvascular endothelial cells, cerebral microvascular endothelial cells, cerebral endothelial cells, human pluripotent stem cells (hPSC), and hPSC-derived endothelial cells. |
| Filed: 12/5/2023 |
| Application Number: US18529927A |
| Tech ID: SD 15162.1 |
| This invention was made with Government support under Contract No. DE-NA0003525 awarded by the United States Department of Energy/National Nuclear Security Administration. The Government has certain rights in the invention. |
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