Supercooling methods for preservation of biological samples
Inventors
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12207647-B2
Publication Date
2025-01-28
Expiration Date
Abstract
The present invention provides methods for cryopreserving a population of cells with improved cell viability. In some aspects, the method comprises contacting a population of cells with a peptoid polymer comprising one or more polar peptoid monomers, e.g., formulated in a cryoprotectant solution, and cooling the population of cells at a temperature of from 0° C. to about −20° C. for a time period of at least about 3 hours to produce a population of supercooled cells. The supercooling methods of the present invention provide excellent post-thaw cell survival and recovery. In certain embodiments, the population of cells is present in a tissue or an organ that is cryopreserved by performing the supercooling methods of the present invention.
Core Innovation
The invention relates to preserving a biological sample or biological macromolecule by contacting the sample or macromolecule with a solution comprising a peptoid polymer or a salt thereof comprising one or more polar peptoid monomers. The peptoid polymer is described as a cryo-protectant that reduces or inhibits ice crystal formation at subzero temperatures, and the preserved biological sample or biological macromolecule is maintained in an unfrozen state.
The method includes cooling the solution to a temperature of from 0°C to about −20°C for a time period of at least about 3 hours to produce a preserved biological sample or preserved biological macromolecule. The disclosed approach addresses limitations of standard cryoprotectant agents by providing an ice-inhibiting preservation strategy under sub-zero supercooling conditions while maintaining the preserved material unfrozen.
The disclosed peptoid polymer embodiments include polar peptoid monomers, including hydroxyl-containing side chains and oligo(ethylene glycol) side-chain chemistries, together with peptoid-peptide hybrid embodiments and specified peptoid polymer salt forms. The disclosure further describes polar and hydrophobic monomer arrangements, repeat length, stereochemical options, and X and Y terminus or covalent cyclized forms.
Claims Coverage
The claim coverage centers on one preservation method using a peptoid polymer, or a salt thereof, comprising one or more polar peptoid monomers to reduce or inhibit ice crystal formation while cooling to 0°C to about −20°C for at least about 3 hours. Across the claim set, there are 4 core inventive features, with dependent claims further refining temperature, time, polymer composition, monomer side-chain chemistry, and salt forms.
Peptoid polymer solution with polar monomers
Contacting a biological sample or biological macromolecule with a solution comprising a peptoid polymer or a salt thereof comprising one or more polar peptoid monomers, wherein the peptoid polymer is present in an amount sufficient to reduce or inhibit ice crystal formation at the temperature.
Cooling to an unfrozen preserved state
Cooling the solution to a temperature of from 0°C to about −20°C for a time period of at least about 3 hours to produce a preserved biological sample or preserved biological macromolecule, wherein the solution comprising the preserved biological sample or biological macromolecule is unfrozen at the temperature.
Polar monomer composition constraints
The peptoid polymer includes at least a specified minimum number of polar peptoid monomers, and the one or more polar peptoid monomers independently include side chains containing a hydroxyl group.
Specified peptoid polymer salt forms
The peptoid polymer salt is chosen from listed counterion salts or combinations thereof, including hydrochloride salt, acetate salt, sulfate salt, phosphate salt, maleate salt, citrate salt, mesylate salt, nitrate salt, tartrate salt, and gluconate salt.
The claims cover preservation of biological samples and biological macromolecules using peptoid polymers, or salts thereof, with polar peptoid monomers to reduce or inhibit ice crystal formation while maintaining an unfrozen state during cooling. Dependent claims further narrow the temperature and time conditions and specify polymer composition features and salt forms.
Stated Advantages
Reduces or inhibits ice crystal formation at the temperature.
Produces a preserved biological sample or preserved biological macromolecule with the solution remaining unfrozen at the temperature.
Preserves a biological sample or biological macromolecule.
Enables preservation across diverse biological samples, including cells, tissues and organs, embryos, sperm/oocytes, and biological macromolecules such as nucleic acids, proteins, and liposomes.
Addresses limitations of standard cryoprotectant agents by providing an ice-inhibiting preservation strategy under sub-zero supercooling conditions.
Documented Applications
Preserving a biological sample or biological macromolecule using a peptoid polymer or a salt thereof comprising one or more polar peptoid monomers in a cooled, unfrozen solution.
Preserving cells, including primary cells and cell lines, by maintaining the preserved solution unfrozen at sub-zero temperatures while reducing or inhibiting ice crystal formation.
Preserving tissues and organs, including bioengineered tissue and organ-related sample types.
Preserving embryos.
Preserving sperm/oocytes.
Preserving biological macromolecules including nucleic acids, proteins, and liposomes.
Assessment of preserved materials using ice detection, thermal characterization, and cell viability assay frameworks.
Interested in licensing this patent?