Acetate-Citrate Clinical Media for Extended Cell Storage
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Solution Overview
Problem
Current clinical media for cell and tissue storage and transport support viability and function only for short periods, often leading to significant cell loss and precipitation issues due to bicarbonate anion reactions with calcium and magnesium salts, and are not compatible with steam sterilization.
Innovation Solution
Development of one-part clinical solutions that utilize acetate and citrate buffers, divalent cations like calcium and magnesium, and glucose to maintain physiological pH and osmolarity, without zwitterionic organic buffers, ensuring stability and prolonged shelf life.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If bicarbonate anions are used as buffering agents in clinical media, then physiological pH buffering is achieved, but carbon salt precipitation occurs with calcium and magnesium salts during heat sterilization and storage
Solution Approach 1:
The patent removes bicarbonate anions from the clinical media formulation to eliminate the precipitation problem. Instead, it uses alternative buffering systems (such as HEPES or other zwitterionic buffers) that do not react with calcium and magnesium salts to form precipitates, thereby extracting the harmful buffering mechanism while preserving the essential pH control function.
Solution Approach 2:
The patent introduces intermediary buffering agents (zwitterionic buffers like HEPES, MOPS, or PIPES) that mediate pH control without forming precipitates with divalent cations. These intermediary substances replace bicarbonate's buffering role while being chemically compatible with calcium and magnesium salts, thus solving the precipitation issue.
2Reliability
If heat sterilization is applied to clinical media containing bicarbonate and calcium/magnesium salts, then sterilization is achieved, but rapid carbonate precipitation occurs
Solution Approach 1:
The patent removes bicarbonate anions from the formulation to prevent their reaction with calcium and magnesium salts during heat sterilization. By eliminating the reactive bicarbonate component, the media can withstand autoclaving without forming carbonate precipitates, thus maintaining solution clarity and compositional stability.
Solution Approach 2:
The patent changes the chemical composition parameters of the buffering system to be heat-stable. By selecting buffers with higher thermal stability and lower reactivity toward divalent cations at elevated temperatures, the media becomes compatible with heat sterilization processes while preventing precipitation.
3Object-generated harmful factors
If two-part kits are used to separate bicarbonate buffer and calcium/magnesium salts, then precipitation is prevented, but manufacturing complexity and mixing errors increase
Solution Approach 1:
The patent merges the previously separated components into a single unified formulation by replacing bicarbonate with heat-stable, precipitation-free buffering agents. This allows all components (including calcium and magnesium salts) to be combined in one stable solution that can be manufactured, stored, and sterilized as a single product without requiring separate containers or mixing steps.
Solution Approach 2:
The patent extracts the problematic bicarbonate component and replaces it with alternative buffers that enable single-part formulation. This elimination of the reactive bicarbonate allows the media to be formulated as a stable one-part product, simplifying the kit structure while maintaining precipitation prevention.
4Device complexity
If zwitterionic organic buffers like HEPES are used to prevent precipitation, then one-part formulation is achieved, but compatibility with cell or tissue culture media is lost
Solution Approach 1:
The patent modifies the buffer selection parameters to identify zwitterionic buffers that are both precipitation-free and compatible with cell culture applications. By carefully selecting buffers with appropriate pKa values, low toxicity, and proven compatibility with mammalian cells, the media achieves both one-part formulation simplicity and broad adaptability for clinical and research uses.
Solution Approach 2:
The patent selects buffering agents that serve multiple functions: they prevent calcium and magnesium precipitation, maintain physiological pH, and are compatible with cell and tissue culture requirements. This multi-functional buffer selection enables the single formulation to be universally applied across different clinical and research applications without requiring separate optimized media.
5Duration of action of stationary object
If current clinical media are used for extended storage, then storage duration is increased, but cell viability and function are significantly lost
Solution Approach 1:
The patent optimizes multiple formulation parameters including buffer type, osmolarity, pH, and additive composition to create a storage environment that maintains cell viability over extended periods. By adjusting these parameters to be more physiologically supportive and stable, the media enables longer storage durations while preserving cell function and reducing stress responses.
Solution Approach 2:
The patent introduces intermediary protective agents and optimized buffer systems that mediate between the storage conditions and living cells. These intermediaries protect cells from stress during extended storage by maintaining stable pH, preventing precipitation, and providing a more physiologically compatible environment, thereby preserving cell viability and function over time.
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 solutions support cell viability and function for extended periods, up to 24-48 hours or more, with reduced precipitation and improved compatibility with steam sterilization, allowing remote formulation and administration without medium replacement.
Implementation Method 1
utilize acetate and citrate buffers, divalent cations like calcium and magnesium, and glucose to maintain physiological pH and osmolarity
Implementation Method 2
utilize acetate and citrate buffers, divalent cations like calcium and magnesium, and glucose to maintain physiological pH and osmolarity
Data Source
AI summary
Some aspects of this disclosure provide clinical media that support viability, re-plating efficiency, and repopulation capacity of cells and tissues during storage for up to 48 hours or longer. The clinical media provided herein are also useful for clinical irrigation. Cell or tissue preparations comprising a cell population or tissue and a clinical medium as provided herein are also provided, as are methods for generating such preparations. Methods for using the clinical media and cell and tissue preparations provided herein, for example, for administering an effective amount of cells or tissue to a subject in need thereof, are also disclosed.