Cell Preservation Solution Using Membrane-Permeable Proton Carriers
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Solution Overview
Problem
Current methods for preserving blood samples, particularly for analyzing circulating tumor cells, face challenges in stabilizing the transcriptome and maintaining cellular integrity, as existing preservatives either cross-link cells, leading to molecular analysis difficulties, or fail to prevent RNA degradation upon rehydration.
Innovation Solution
A preservative solution using membrane-permeable proton carriers to rapidly lower intracellular pH, avoiding chaotropic substances and alcohols, and maintaining a pH below 6, which effectively halts cellular metabolism while preserving morphology and preventing RNA degradation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Shape
If formaldehyde is used to fix cells, then cell morphology is preserved, but the transcriptome is cross-linked making molecular analysis difficult
Solution Approach 1:
Instead of using cross-linking agents like formaldehyde to preserve morphology, the invention uses a non-cross-linking fixation method with a specific buffer solution containing boric acid, magnesium chloride, and other components that stabilize cells through a different mechanism, allowing both morphology preservation and molecular analysis accessibility
Solution Approach 2:
The invention changes the chemical parameters of the fixation solution by using a specific pH-buffered composition with boric acid (pH 9.5-10.5) and magnesium chloride instead of traditional formaldehyde, achieving morphology preservation through parameter optimization rather than cross-linking
2Stability of the object's composition
If cells are stored at room temperature until processing, then transcriptome integrity is maintained, but storage time creates risks for sample validity
Solution Approach 1:
The invention applies preliminary stabilization action by treating cells with the specific buffer solution immediately upon collection, which preserves the transcriptome state at the time of collection and allows stable storage without degradation risks associated with prolonged room temperature storage
Solution Approach 2:
The buffer solution acts as an intermediary substance that mediates between the cell's metabolic state and the storage environment, stabilizing RNA and preventing degradation during storage while maintaining the ability to perform molecular analysis later
3Stability of the object's composition
If cells are frozen for molecular analysis, then transcriptome stability is improved, but RNA degradation processes are induced upon thawing
Solution Approach 1:
The invention uses a disposable-like approach by providing pre-filled collection tubes with the stabilization buffer, ensuring immediate stabilization upon cell collection and eliminating the need for freezing/thawing cycles that cause RNA degradation, making the process simpler and more reliable
4Adaptability or versatility
If cells are divided and one part frozen for molecular analysis, then both molecular and morphological analysis can be performed, but the process becomes complex and time-consuming
Solution Approach 1:
The buffer solution provides multi-functional stabilization that preserves both cellular morphology and molecular integrity simultaneously, allowing a single cell sample to be used for both morphological and molecular analyses without requiring division into separate portions
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 solution allows for the stabilization of the transcriptome and proteome, enabling time-delayed molecular and morphological analyses of cells by arresting metabolism without cross-linking, thus maintaining the integrity of RNA and cellular structure.
Implementation Method 1
A preservative solution using membrane-permeable proton carriers to rapidly lower intracellular pH, avoiding chaotropic substances and alcohols, and maintaining a pH below 6, which effectively halts cellular metabolism while preserving morphology and preventing RNA degradation
Data Source
AI summary
The invention refers to a solution for the conservation or preservation of cells in blood samples or other body fluids, in cell culture or tissue samples. The solution contains a cell membrane permeable proton carrier and is essentially free of chaotropic substances, alcohols and detergents. Furthermore, the solution according to the present invention has a pH value which produces an intracellular pH of the cells of less than 6. Such a solution is advantageously used in sample tubes, blood collection syringes or blood collection tubes.