Cryogenically Arrayable Stable Transfection Workflow
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for gene editing in cells are time-consuming and require extensive preparation, including cell expansion and centrifugation, which slows down the process of editing multiple cells and cell lines simultaneously while maintaining efficiency and safety.
Innovation Solution
The Cryogenically Arrayable Stable Transfection (CAST) method involves thawing cryopreserved cells directly into a transfection workflow without the need for expansion or centrifugation, allowing for immediate contact with genetic material and a cell manipulation solution to introduce the genetic material into the cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional transfection methods are used with cell expansion and centrifugation steps, then cell viability and transfection efficiency are maintained, but processing time increases significantly
Solution Approach 1:
Cells are cryopreserved in a ready-to-use state with freezing media already in place, eliminating the need for post-thaw expansion. The cells are prepared in advance through cryopreservation, allowing immediate transfection upon thawing without requiring time-consuming expansion steps.
Solution Approach 2:
The invention extracts and removes the time-consuming cell expansion step from the traditional transfection workflow. By using cryopreserved cells that can be directly transfected after thawing, the expansion phase is completely eliminated, reducing processing time while maintaining transfection efficiency.
2Loss of time
If cells are cryopreserved and thawed without centrifugation, then processing time is reduced, but cell viability may be compromised
Solution Approach 1:
The freezing media serves dual purposes: it maintains cells in a viable state during cryopreservation and enables direct transfection after thawing without requiring centrifugation or expansion steps. The media itself facilitates the transfection process, eliminating the need for additional cell handling steps.
Solution Approach 2:
The invention changes the physical state and composition parameters of the cell suspension by using freezing media that allows direct transfection. The media's chemical composition and physical properties are optimized to enable immediate transfection upon thawing, maintaining cell viability without centrifugation.
3Productivity
If multiple cell lines are processed simultaneously, then productivity increases, but maintaining consistent transfection efficiency becomes more difficult
Solution Approach 1:
The cryopreservation and transfection protocol is designed to be universally applicable across multiple cell lines and types. The same freezing media composition and transfection protocol can be used for different cell lines, enabling simultaneous processing while maintaining consistent transfection efficiency through standardized procedures.
Solution Approach 2:
The invention segments the transfection process into discrete, standardized steps that can be applied to multiple cell lines simultaneously. By using arrayed formats and standardized protocols for each cell line, consistent transfection efficiency is maintained across parallel processing of multiple cell types.
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
CAST enables accelerated gene editing by eliminating the need for cell expansion and centrifugation, thereby reducing processing time and maintaining high editing efficiency across multiple cell types and cell lines.
Implementation Method 1
cells are cryopreserved within a freezing media in a cell container
Implementation Method 2
thawing the cells in the cell container
Data Source
AI summary
Provided herein systems and methods for processing cryopreserved cells without an expansion phase after thawing, including transfecting cells for gene editing purposes and cryopreserving transfected cells. Systems and methods for high-throughput processing of frozen cells are also provided. The systems and methods provided can be used to modify cells, including gene editing, in cell arrays.