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US 12,540,938 B1Granted
Real-time patch-clamp sensing of transport across a barrier tissue
AssigneeNational Technology & Engineering Solutions of Sandia, LLC
InventorsRamdane Abdallah Harouaka, Michael Bartsch, Jennifer Lauren Schwedler, Brooke Nicole Harmon
Life of this patent
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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.
Claims20
A method comprising:
- culturing a plurality of barrier cells on a first surface of a substrate and within a first chamber, wherein the substrate comprises one or more pores disposed within the substrate, and wherein at least one pore is a micropore or a nanopore, thereby forming a seal between at least one pore and at least one barrier cell;
- adding an agent to the first chamber;
- and
- measuring one or more electrical signals within the first chamber or within a second chamber in fluidic communication with the at least one pore, wherein the one or more electrical signals is indicative of transport or lack of transport of the agent through the at least one barrier cell;
- wherein the plurality of barrier cells comprises a layer of cells wherein the barrier cell remains functionally active and intact during the measuring step;
- wherein the electrical signals are transmitted in real-time as the transport or lack of transport of the agent through the at least one barrier cell is detected;
- wherein the plurality of barrier cells comprises a co-culture of a barrier cell selected from one or more of the group consisting of:
- brain microvascular endothelial cells, cerebral microvascular endothelial cells, cerebral endothelial cells, human pluripotent stem cells (hPSC), and hPSC-derived endothelial cells.
5 more elements
A method comprising:
- culturing a plurality of barrier cells on a first surface of a substrate and within a first chamber, wherein the substrate comprises one or more pores disposed within the substrate, and wherein at least one pore is a micropore or a nanopore, thereby forming a seal between at least one pore and at least one barrier cell;
- adding an agent to the first chamber;
- and
- measuring one or more electrical signals within the first chamber or within a second chamber in fluidic communication with the at least one pore, wherein the one or more electrical signals is indicative of transport or lack of transport of the agent through the at least one barrier cell;
- wherein the electrical signals are transmitted in real-time as detections are made of agents transported through vesicles formed on one surface of the at least one barrier cell and to an opposite side of the at least one barrier cell;
- wherein the seal is gigaohm seal and the measuring step is completed without disturbing barrier function or cell viability
- wherein the plurality of barrier cells comprises a confluent layer of barrier cells having tight junctions and/or adherens junctions.
4 more elements
Dependencies are inferred from the claim text.