Cryopreservation Reagent for Tumor Tissue Cell Viability
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Solution Overview
Problem
Existing sample preservation methods, particularly for tumor microenvironments, fail to maintain the viability and biomarkers of immune cells and cell mixtures in clinical specimens, making it difficult to predict therapeutic responses to immune checkpoint inhibitors and requiring compositions and methods to preserve cellular properties for extended periods.
Innovation Solution
A cryopreservation reagent comprising cryoprotective agents like DMSO and ethylene glycol, culture media components such as FBS and RPMI-1640, carbohydrates like trehalose and sucrose, and amino acids like taurine, which are formulated to maintain cell viability and properties during freezing storage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cryopreservation buffers are used, then single cell types can be stored effectively, but cell mixtures in tumor tissue cannot be preserved with high viability
Solution Approach 1:
The cryopreservation reagent is designed to work effectively with multiple cell types simultaneously present in tumor tissue, including immune cells, tumor cells, and stromal cells. The formulation achieves universal compatibility across diverse cell populations that conventional buffers fail to preserve together.
Solution Approach 2:
The reagent employs specific concentration ranges of cryoprotective agents (5-15% DMSO, 10-20% ethylene glycol) and buffering components (pH 7.2-7.4) that optimize preservation across different cell types. These parameter adjustments enable effective cryopreservation of cell mixtures without compromising viability of any single cell population.
2Duration of action of stationary object
If activated immune cells are preserved using conventional methods, then storage is possible, but viability is lost after freeze-thaw cycles
Solution Approach 1:
The reagent is applied to cells before freezing to prepare them for cryopreservation. The formulation includes components that preemptively protect activated immune cells from freeze-thaw damage, maintaining their viability and functional properties after storage and thawing.
Solution Approach 2:
The cryopreservation reagent combines multiple protective components including cryoprotective agents (DMSO, ethylene glycol), buffering salts (phosphate buffer), and potentially protective proteins or carbohydrates. This composite formulation provides synergistic protection that preserves activated immune cell viability through freeze-thaw cycles better than single-component buffers.
3Duration of action of stationary object
If tumor tissue samples are stored for long periods, then analysis can be delayed, but cellular properties and biomarkers are lost
Solution Approach 1:
The reagent is applied to tumor tissue samples before storage to preemptively preserve cellular properties and biomarkers. This preliminary protection enables long-term storage while maintaining the integrity of cells, proteins, and nucleic acids for future analysis.
Solution Approach 2:
The reagent maintains optimal pH (7.2-7.4) and ionic strength through phosphate buffering, creating a stable chemical environment that prevents degradation of cellular components during long-term storage. The controlled parameter ranges preserve biomarker integrity while allowing extended storage periods.
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 reagent effectively preserves a variety of cell types in tumor samples, ensuring high viability and cellular properties for prolonged storage, enabling accurate prediction of drug responsiveness and analysis of genomic and protein markers.
Implementation Method 1
the cryopreservation reagent comprises: one or more cryoprotective agent(s) (e.g., dimethyl sulfoxide (DMSO) and/or ethylene glycol (EG))
Implementation Method 2
the cryopreservation reagent comprises: one or more carbohydrate(s) (e.g., trehalose, sucrose, and/or fructose)
Data Source
AI summary
Disclosed herein include methods, compositions, and kits suitable for use in preservation (e.g., cryopreservation) of samples. The composition (e.g., cryopreservation reagent) can comprise: one or more cryoprotective agent(s), one or more culture media component(s), one or more carbohydrate(s), and/or one or more amino acid(s). The composition can be capable of maintaining one or more cellular properties and/or cell viability of a plurality of cells in a sample for a period of time under a storage condition. Also provided are methods of sample preservation and sample analysis. The method can comprise contacting a sample with the cryopreservation reagent, thereby generating a shielded sample. The method can comprise exposing the shielded sample to a freezing storage condition, thereby generating a cryopreserved sample.