Continuous-Flow Cell Membrane Delivery With Reversible Permeabilization
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Solution Overview
Problem
Existing methods face challenges in delivering molecules across cell membranes, particularly for larger, charged, or complex molecules, often resulting in irreversible permeabilization and cell viability issues.
Innovation Solution
A continuous flow fluidic system using a microfluidic device with controlled mixing and permeabilization, followed by a stop solution to maintain cell viability, enabling reversible permeabilization and efficient delivery of payloads across cell membranes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If batch processes are used for payload delivery, then cell processing can be performed, but the throughput is limited and processing time is excessive
Solution Approach 1:
The system pre-prepares cells in suspension and delivers payloads before final cell recovery, enabling continuous processing without waiting for each batch to complete all steps. This preliminary arrangement of materials and methods allows high-throughput continuous operation while maintaining effective payload delivery to cells.
2Productivity
If permeabilization agents are used to deliver payloads across cell membranes, then delivery efficiency is improved, but cell viability is reduced due to irreversible permeabilization
Solution Approach 1:
The system uses periodic or controlled exposure to permeabilization agents rather than continuous exposure. By controlling the timing and duration of permeabilization agent contact with cells in continuous flow, the system achieves effective payload delivery while allowing cells to recover, thereby maintaining cell viability through cyclical rather than sustained permeabilization.
Solution Approach 2:
The system uses an intermediary controlled environment (continuous flow system with defined residence time) that mediates between the permeabilization agent and cells. This intermediary system allows precise control of exposure duration and conditions, enabling payload delivery while protecting cell viability through controlled interaction parameters.
3Productivity
If continuous flow system is implemented, then throughput is improved, but system complexity increases
Solution Approach 1:
The continuous flow system integrates multiple functions into a single unified platform: cell suspension delivery, payload delivery, controlled permeabilization, and cell recovery all occur in sequence within one system. This multi-functionality achieves high throughput while avoiding the need for multiple separate batch processing systems, thereby managing complexity through integration rather than proliferation of separate components.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The system achieves consistent, high-throughput delivery of various payloads to cells while maintaining cell viability, allowing for therapeutic applications and scalable processes.
Implementation Method 1
a delivery solution that causes the population of cells to experience permeabilization and delivers the delivery payload across membranes of the population of cells
Implementation Method 2
a stop solution that causes the population of cells to stop experiencing permeabilization or experience a reduction in permeabilization
Data Source
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AI summary
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.