Layered Cell Sheet Production via Centrifugal Force and Temperature Control
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for layering cell sheets are time-consuming and struggle to maintain adhesiveness to temperature-responsive culture dishes, making it difficult to efficiently produce and transfer three-dimensional tissues.
Innovation Solution
A method involving the application of a centrifugal force to cell sheets on a temperature-responsive culture surface within a specific temperature range, allowing for rapid layering and non-invasive transfer of cell sheets by controlling the temperature and centrifugal force parameters.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a centrifugal force is applied to layer cell sheets at room temperature, then the layering time is shortened, but the cell sheets cannot be non-invasively obtained and transferred
Solution Approach 1:
The patent applies centrifugal force at physiological temperature (37°C) rather than room temperature, changing the temperature parameter to enable both rapid layering and subsequent non-invasive transfer. This parameter change allows the cell sheets to maintain their adhesiveness to the temperature-responsive culture dish during centrifugation while still enabling easy release and transfer after layering is complete.
2Reliability
If conventional layering methods are used without centrifugal force, then cell sheet adhesiveness is maintained, but the layering process takes 30-60 minutes
Solution Approach 1:
The patent uses periodic centrifugal force application (e.g., 5 minutes at 50×g) to accelerate the layering process while maintaining cell sheet adhesiveness. The centrifugal force is applied in a controlled, periodic manner that promotes rapid cell sheet attachment without compromising the biological integrity or adhesiveness of the cells, achieving both speed and reliability.
3Productivity
If centrifugal force is applied at room temperature for rapid layering, then productivity increases, but the morphology of cell sheets deteriorates
Solution Approach 1:
The patent changes the temperature parameter from room temperature to physiological temperature (37°C) during centrifugal force application. This parameter change maintains cell sheet morphology by keeping the cells in a physiologically appropriate environment, preventing deformation or damage that would occur at lower temperatures, while still achieving rapid layering through the centrifugal force.
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
This method significantly shortens the layering time, maintains the morphology of the cell sheets, and enables easy transfer of the layered cell sheets to target surfaces, enhancing the efficiency of tissue production and transplantation.
Implementation Method 1
lowering the temperature to 20° C., which is less than 32° C. of a lower critical solution temperature (LCST) of PIPAAm
Implementation Method 2
a technique of shorting the layering time by applying a centrifugal force to the layered cell sheet by means of a centrifuge for plates
Data Source
AI summary
The present invention provides a method for swiftly producing a layered cell sheet that is non-invasively obtained and is utilizable for transplantation, etc., the method including (1) a step of applying a centrifugal force to a first cell sheet on a temperature-responsive culture surface for a predetermined time in a temperature range from a lower critical solution temperature of the temperature-responsive culture surface to 45° C., (2) a step of further placing a second cell sheet on the first cell sheet, and (3) a step of applying a centrifugal force to the first cell sheet and the second cell sheet on the temperature-responsive culture surface for a predetermined time in the temperature range from the lower critical solution temperature to 45° C.; and also provides a layered cell sheet obtained by the method.