Cell Patch Cryopreservation Composition for Structure and Viability
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing cell patch cryopreservation methods fail to maintain structural integrity and cellular viability due to disruption of cell-to-cell connections and low survival rates, necessitating a solution that ensures high cell density and intact structure during cryopreservation.
Innovation Solution
A cell patch cryopreservation solution comprising a cryoprotectant, buffer, reducing agent, calcium channel inhibitor, and apoptosis inhibitor, with specific concentrations of components like DMSO, HEPES buffer, VC, verapamil, and Caspase inhibitor, is used to preserve cell patches.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional cell cryopreservation solutions are used, then cell survival rate can be maintained, but cell patch structure is disrupted and cell-to-cell connections are broken
Solution Approach 1:
The patent modifies the chemical composition parameters of the cryopreservation solution by incorporating specific concentrations of calcium channel inhibitors (5-500 μM), apoptosis inhibitors (1-50 μM), and reducing agents (0.1-10 mM), which fundamentally changes how cells undergo cryopreservation while maintaining structural integrity
Solution Approach 2:
The invention creates a composite cryopreservation solution containing multiple functional components working synergistically: cryoprotectants (DMSO, sucrose, glycerol), buffers (HEPES, phosphate buffer), reducing agents (VC, glutathione, allopurinol), calcium channel inhibitors, and apoptosis inhibitors, forming a complex protective matrix that simultaneously preserves cell viability and structural integrity
2Ease of operation
If cell patches are stored in ready-to-use manner, then immediate application is possible, but storage time is limited to 24 hours and requires advance appointment scheduling
Solution Approach 1:
The patent enables preliminary preparation and storage of cell patches by developing a cryopreservation solution that maintains cell viability and structural integrity over extended periods, allowing patches to be prepared in advance and stored without degradation, thus eliminating the 24-hour storage limitation
Solution Approach 2:
By changing the chemical environment through the addition of specific inhibitors and protectants, the patent extends the temporal stability of cell patches, transforming them from short-term storage products to long-term shelf-stable products
3Reliability
If classical safety tests are performed before use, then patient safety is ensured, but results cannot be obtained due to short window period and require rapid detection methods increasing cost and risk
Solution Approach 1:
The patent allows safety testing to be performed in advance on stored cell patches before patient transplantation, eliminating the time window constraint by enabling preliminary preparation and storage of tested samples, thus ensuring patient safety without rushing the detection process
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 solution effectively maintains the structural integrity and achieves a cell survival rate of over 80% after cryopreservation and resuscitation, addressing the limitations of traditional methods.
Implementation Method 1
a cryoprotectant... the cryoprotectant is at least one selected from a group consisting of DMSO with a volume concentration of 2% to 10%, sucrose with a mass concentration of 2 g/100 mL to 10 g/100 mL, ethylene glycol with a volume concentration of 1% to 5%, cross-linked polyvinylpyrrolidone with a mass concentration of 2 g/100 mL to 10 g/100 mL, propylene glycol with a volume concentration of 2% to 10%, trehalose with a mass concentration of 2 g/100 mL to 10 g/100 mL, methylcellulose with a mass concentration of 0.05 g/100 mL to 0.5 g/100 mL, glycerol with a volume concentration of 5% to 15% based on the whole cell patch cryopreservation solution
Implementation Method 2
a reducing agent... the reducing agent is at least one selected from a group consisting of VC with a mass concentration of 1 to 100 μg/mL, reduced glutathione with a molar concentration of 0.3 to 30 mM, allopurinol with a molar concentration of 0.1 to 10 mM, adenylate with a molar concentration of 0.5 to 50 mM, based on the whole cell patch cryopreservation solution
Implementation Method 3
a calcium channel inhibitor... the calcium channel inhibitor is at least one selected from a group consisting of anipamil, falipamil, devapamil, gallopamil, tiapamil and verapamil. Optionally, a molar concentration of the calcium channel inhibitor is 5 to 500 M based on the whole cell patch cryopreservation solution
Implementation Method 4
an apoptosis inhibitor... the apoptosis inhibitor is at least one selected from a group consisting of a P53 inhibitor and a Caspase inhibitor. Optionally, a molar concentration of the apoptosis inhibitor is 1 to 50 M based on the whole cell patch cryopreservation solution
Implementation Method 5
a buffer... the buffer comprises one of a HEPES buffer system, a phosphate buffer system, a MOPS buffer system, a PIPES buffer system
Data Source
AI summary
The present disclosure provides a cell patch cryopreservation solution, which includes: a cryoprotectant, a buffer, a reducing agent, a calcium channel inhibitor and an apoptosis inhibitor. The present disclosure also provides a method of cryopreserving a cell patch by using the aforementioned cell patch cryopreservation solution. The cell patch cryopreservation solution and the method of cryopreserving a cell patch provided by the present disclosure can maintain the structural integrity and the cell activity of the cell patch.