Immobilized EV-Coated Surfaces for Reproducible In Vitro Assays
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Solution Overview
Problem
The purification of extracellular vesicles (EVs) is costly, tedious, and lacks reproducibility, with existing methods leading to quality loss and contamination issues, and their storage is limited, making them unsuitable for reliable in vitro assays.
Innovation Solution
EVs are immobilized on a sterile surface by drying, allowing them to be frozen and stored without buffer components, maintaining their activity for extended periods, and enabling the creation of an EV library for parallel testing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If EVs are stored in solution in the fridge, then they remain accessible for use, but their shelf-life is limited and quality declines over days
Solution Approach 1:
The patent changes the physical state of EVs from dissolved in solution to immobilized on a surface. This parameter change allows EVs to be stored for extended periods (weeks to months) without quality degradation, as the immobilized EVs are not subjected to buffer components or freezing conditions that cause quality loss in solution-based storage
Solution Approach 2:
The patent extracts EVs from solution by immobilizing them on a surface through drying. This removes EVs from the problematic solution environment containing buffer components that limit shelf-life and cause quality degradation during storage
2Duration of action of stationary object
If EVs are frozen in solution with buffer components, then they can be stored long-term, but the buffer components may not be compatible with in vitro assays and EV quality is lost
Solution Approach 1:
The patent extracts EVs from solution by immobilizing them on a surface through drying, removing them from the problematic buffer environment before storage. This eliminates the harmful interaction between buffer components and EVs during long-term storage
Solution Approach 2:
The patent performs preliminary drying to immobilize EVs on the surface before storage. This preliminary action removes the solution and buffer components that would cause incompatibility issues and quality loss during subsequent long-term storage or freezing
3Quantity of substance
If EVs are purified using conventional methods, then EV preparations can be obtained, but the process is costly, tedious, and lacks reproducibility
Solution Approach 1:
The patent uses a self-service approach where EVs in the supernatant are directly immobilized on cell culture surfaces through simple drying without requiring complex purification equipment or multiple centrifugation steps. This dramatically simplifies the manufacturing process while maintaining EV quality
Solution Approach 2:
The patent extracts the essential EV-containing supernatant from cell cultures and directly immobilizes it on surfaces, taking out the need for complex purification steps while retaining the functional EV preparations
4Ease of operation
If EVs are used in in vitro assays with buffer components, then assays can be performed, but buffer components and impurities may interfere with cell responses
Solution Approach 1:
The patent extracts EVs from buffer solution by immobilizing them on a surface through drying. This removes the harmful buffer components and impurities that would interfere with in vitro assays, leaving only the EVs available for cell interactions
Solution Approach 2:
The patent uses the cell culture surface as an intermediary medium to deliver EVs to cells without requiring buffer components. The surface acts as a mediator that presents EVs to cells in a clean, interference-free environment
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 immobilization method provides stable, reproducible EVs that can be stored for weeks, facilitating efficient in vitro assays by eliminating buffer-related issues and allowing simultaneous testing of various EV sources and concentrations.
Implementation Method 1
a solution comprising EVs may be dried on a cell culture surface to obtain a surface being coated with immobilized EVs
Data Source
AI summary
Isolation of extracellular vesicles (EVs) is a tedious and costly process and EVs can often not be stored well. The application relates to a sterile surface suitable for culturing cells, wherein the surface is coated with extracellular vesicles (EVs7) immobilized at one or more predetermined position(s). It has been surprisingly found that extracellular vesicles (EVs) can be immobilized on a sterile surface, for example by drying. Additionally, a device for culturing cells comprising said surface further comprising a removable covering to preserve sterility of said surface is disclosed. Furthermore, a method of preparing said sterile surface is disclosed. Finally, the application also discloses methods of monitoring cell differentiation, apoptosis, cell migration, proliferation, transcriptome, proteome, and/or viability of cells (recipient cells'), wherein said recipient cells are added to said EV-coated surface and then monitored.