Mechanical Transfection Device Using Microporous Membrane
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Solution Overview
Problem
Current transfection methods often damage cells and are inefficient for delivering large cargo molecules across the cellular membrane, particularly for materials larger than 1 μm, as they require significant disruption of the cellular membrane, limiting the throughput and effectiveness of transfection.
Innovation Solution
A transfection device utilizing a deformable fluid reservoir coupled to a container via a microporous membrane, where a macrostructure is delivered under pressure to cells, facilitating its internalization via active endocytosis, with a pneumatic or mechanical actuator exerting force to move the macrostructure through the membrane pores, maintaining low cell damage and high transfection efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If significant disruption of cellular membrane is applied to deliver large cargo material, then delivery efficiency is improved, but cell damage increases
Solution Approach 1:
The patent employs a porous membrane with controlled pore sizes (0.2-10 μm) to facilitate the delivery of large cargo materials into cells. The porous structure allows cargo to pass through while maintaining cell integrity, resolving the contradiction between delivery efficiency and cell damage by providing a physical pathway that does not require severe membrane disruption
Solution Approach 2:
The porous membrane acts as an intermediary element between the cargo material and the cells. It enables controlled interaction by allowing cargo to pass through its pores while protecting the cell from direct exposure to harsh delivery conditions, thus improving delivery efficiency without causing significant cell damage
2Quantity of substance
If poration is used to transfect cells, then material delivery is achieved, but the amount of material delivered is significantly reduced for large molecules
Solution Approach 1:
The porous membrane provides continuous physical pathways for cargo delivery, eliminating the need for transient poration. This allows sustained and efficient delivery of large cargo materials (proteins, viruses, organelles) without the limitations of poration-based methods, thereby increasing both the quantity delivered and transfection efficiency
3Quantity of substance
If shockwave or pressure is applied to accelerate micro projectiles into cells, then delivery of large molecules is guaranteed, but cell damage is significantly increased
Solution Approach 1:
The patent applies controlled hydrostatic pressure to drive cargo through the porous membrane into cells. This hydraulic approach delivers large molecules effectively while maintaining controlled, non-traumatic pressure levels that avoid the severe cell damage associated with shockwave methods
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 device achieves high transfection efficiency with minimal cell damage, capable of delivering macrostructures like mitochondria and nucleic acid lipoplexes, demonstrating successful colony formation and GFP expression, outperforming traditional methods in terms of throughput and cell viability.
Implementation Method 1
a macrostructure is delivered under pressure from a deformable fluid reservoir to cells contained in a container
Implementation Method 2
the deformable fluid reservoir is coupled to the container via a microporous membrane having a pore size suitable to deliver the macrostructure to the cell
Implementation Method 3
which is thought to internalize the macrostructure via active endocytosis
Data Source
AI summary
A transfection device suitable for delivery of various macrostructures (e.g., mitochondria, bacteria, liposomes) is described and uses mechanical force to thereby induce active endocytosis in a target cell. Contemplated devices are able to achieve high throughput of transfected cells that remain viable and are capable of producing colonies.