Payload delivery across cell membranes using continuous flow fluidic system
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Publication Number
US-12104166-B2
Publication Date
2024-10-01
Expiration Date
Abstract
Methods, systems, processes, and apparatuses are provided for delivery across cell membranes. In one aspect, an apparatus includes a substrate including a mixing channel, a process chamber, and a dilution channel to perform delivery of the payload to across the cell membranes. In another aspect, a system includes reservoirs for a cell suspension, a delivery solution, and a stop solution connected to a pump. The system further includes an agitator, a heater, a temperature controller, and a controller to operate the system. In yet another aspect, cells in suspension are mixed with a delivery solution in a microfluidic mixing chip. The delivery solution includes a permeabilization agent to cause permeabilization of the cells, allowing delivery of a payload from the delivery solution to the cells across the cell membranes.
Core Innovation
The invention relates to a method of delivering a payload across a cell membrane. The method mixes a population of cells in a suspension and a delivery payload in a delivery solution within at least one microfluidic channel contained in a substrate with microfluidic channels, so that the mixing occurs within the microfluidic channels during delivery across membranes.
A delivery solution is used that causes the population of cells to experience permeabilization, while the delivery payload is delivered across membranes of the population of cells. The delivery solution includes an alcohol, and the mixed cells and delivery payload are subsequently exposed to a stop solution introduced after mixing.
The stop solution is introduced subsequent to mixing such that it causes the population of cells to stop experiencing permeabilization. At least a portion of the population of cells that stop experiencing permeabilization are viable, and the method includes analyzing the at least a portion of the population of cells that stop experiencing permeabilization and are viable.
Claims Coverage
The independent claim covers a microfluidic mixing-based payload delivery method in which an alcohol-containing delivery solution permeabilizes cells for delivery, and a subsequent stop solution halts permeabilization so viable cells can be analyzed. The independent claim includes five inventive features.
Microfluidic mixing within microfluidic channels
Mixing a population of cells in a suspension and a delivery payload in a delivery solution, with the mixing performed using a substrate containing microfluidic channels and occurring within at least one of the microfluidic channels.
Non-adherent cell population including PBMCs, immune cells, or T lymphocytes
The population of cells includes a population of non-adherent cells, wherein the non-adherent cells comprise peripheral blood mononuclear cells (PBMCs), immune cells, or T lymphocytes.
Alcohol-containing permeabilization delivery across cell membranes
The delivery solution causes the population of cells to experience permeabilization and the delivery payload is delivered across membranes of the population of cells, and the delivery solution includes an alcohol.
Stop solution halts permeabilization after mixing to preserve viable cells
Introducing a stop solution to the population of cells and the delivery payload subsequent to mixing such that the stop solution causes the population of cells to stop experiencing permeabilization; wherein at least a portion of the population of cells that stop experiencing permeabilization are viable.
Analyzing viable cells after stopping permeabilization
Analyzing the at least a portion of the population of cells that stop experiencing permeabilization and are viable.
Overall, the claim coverage centers on a microfluidic-channel mixing approach for delivering a payload across cell membranes using an alcohol-containing permeabilization delivery solution, followed by introducing a stop solution after mixing to halt permeabilization while retaining viability, and analyzing the viable cells after stopping permeabilization.
Stated Advantages
Enables at least a portion of cells that stop experiencing permeabilization to be viable for analysis.
Allows analysis of cells after permeabilization is halted using the stop solution.
Documented Applications
No documented applications found
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