Responsive cell culture hydrogel
Inventors
Schmidt, Daniel • Reynaud, Emmanuelle • Gaines, Peter
Assignees
University of Massachusetts Lowell
Publication Number
US-9506029-B2
Publication Date
2016-11-29
Expiration Date
2031-12-30
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Abstract
The present invention provides devices, compositions and methods for maintaining conditions in a cell culture and for measurement of conditions in the cell culture. In particular, the invention provides hydrogel materials, apparatus and methods for several non-invasive techniques of maintaining optimal or near-optimal nutrient and pH levels in cell cultures.
Core Innovation
The invention provides devices, compositions, and methods for maintaining and measuring conditions in cell cultures using responsive hydrogel materials. These hydrogels are based on synthetic polymers and are notable for their high water content and biocompatibility, enabling several non-invasive techniques to maintain optimal or near-optimal nutrient and pH levels in cell cultures. The system can incorporate pH-sensitive hydrogels that release desired nutrients or absorb components from the cell culture in response to decreasing pH.
Specifically, the hydrogels are constructed from cross-linked polymer networks formed from a pH-sensitive precursor and a linker or cross-linker, and are loaded with one or more agents such as pH-sensitive dyes, glucose, amino acids, growth factors, or pH-regulating agents and buffers. These hydrogels are engineered to swell or deswell in response to changes in environmental pH, resulting in controlled delivery or uptake of these agents. The inclusion of pH-sensitive dyes enables both qualitative visual monitoring and quantitative measurement of pH, allowing for remote or automated broadcasting of cell culture health information.
The problem addressed by the invention is the need for an accurate, automated, and non-invasive system for maintaining cell cultures at optimal conditions of pH and nutrient concentration. Traditional manual processes may lead to periods of suboptimal conditions due to limited intervention frequency, which can result in cell distress, death, or altered characteristics. The invention further provides methods to reduce contamination risks by minimizing direct human interaction and extends cell viability, especially in small-scale cultures often used in high-throughput drug screening.
Claims Coverage
The patent comprises one independent composition claim and several method claims focusing on inventive features related to responsive hydrogel polymers for condition-responsive agent delivery and monitoring in cell cultures.
pH-responsive hydrogel polymer composition for cell culture
A hydrogel polymer comprising: - A pH-sensitive precursor with more than two amine hydrogens per molecule (or at least two amine hydrogens per molecule) - A linker with at least two epoxy groups per molecule (or more than two epoxy groups per molecule) - One or more agents chosen from pH-sensitive dye, nutrient, amino acid, growth factor, cell media pH-regulating agent, and mixtures thereof - The pH-sensitive precursor and linker are covalently attached - The dried hydrogel network exhibits swelling in aqueous environments, with defined quantitative measures for swelling at different pH values (degree of swelling at pH 2 between 10% and 200%; difference between swelling at pH 2 and at pH 9 between 3% and 190%; ratio of swelling at pH 2 to pH 9 between 1.2 and 20.8) - The release of the agents is responsive to changes in cell culture pH
Method for delivering agents to cells using hydrogel polymer
A process including: 1. Adding the hydrogel polymer composition to media containing cells, where the hydrogel is capable of releasing one or more agents. 2. Culturing the cells so that the agents are released into the media and delivered to the cells.
Method for maintaining optimal cell culture pH using hydrogel polymer
A method comprising: 1. Adding the hydrogel polymer to the cell culture, where the agent includes a pH-regulating agent. 2. Culturing cells so that the pH-regulating agent is released in response to a change in pH, maintaining optimal pH conditions within the culture.
Method for maintaining optimal glucose level in cell culture using hydrogel polymer
A method comprising: 1. Adding the hydrogel polymer to the cell culture, loaded with glucose as an agent. 2. Culturing the cell culture so that glucose is released into the culture in response to pH changes, thereby maintaining an optimal glucose level.
Method for maintaining optimal L-glutamine level in cell culture using hydrogel polymer
A method comprising: 1. Adding the hydrogel polymer containing L-glutamine (free or as a dipeptide) to the cell culture. 2. Culturing cells so that L-glutamine is released in response to a change in pH, maintaining optimal L-glutamine levels.
Collectively, these inventive features protect the use of pH-responsive hydrogel compositions for triggered delivery and monitoring of agents in cell cultures, based on responsive polymeric materials defined by their precursor and linker functionalities and swelling/response behaviors.
Stated Advantages
The hydrogels can be produced easily and rapidly from inexpensive, commercially available materials.
The invention reduces the chances of contamination and cell distress or death from nutrient depletion or suboptimal conditions by enabling non-invasive, automated maintenance.
Maintenance of optimal conditions may extend the viability of cells in small volumes, useful for drug screening and for culturing cell types prone to differentiation.
By optimizing cell growth and reducing cell death, the technology allows scientists to accelerate studies, companies to move products faster to market, and provides more reliable experimental results.
The system eliminates the need for direct human intervention, reducing labor costs and allowing personnel to spend less time managing cell cultures.
The hydrogels can be sterilized and stored, feature high mechanical strength, and are biocompatible with low cytotoxicity.
The responsive hydrogels can be tuned for specific cell culture needs through customizable composition and properties.
Non-invasive real-time optical monitoring using pH-sensitive dyes enables continual assessment of cell culture conditions without risking contamination.
Documented Applications
Automatically maintaining optimal pH and nutrient (e.g., glucose, L-glutamine) levels in cell cultures, especially in small-volume formats such as 96-well plates.
Delivering one or more agents (nutrients, amino acids, growth factors, pH-regulating agents) to cells in a controlled, pH-responsive manner during cell culture.
Non-invasive, real-time monitoring of cell culture pH via visual or optical measurement using hydrogels containing pH-sensitive dyes.
Reducing cell culture contamination risk and associated labor in both research and industrial cell culture operations.
Extending the viability of cultured cells and maintaining their characteristics for applications such as drug screening and culturing cells prone to differentiation.
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