Abstract: The present disclosure relates to devices and methods for the storage of material at cryogenic temperatures. Such devices may be useful for storing materials in the vapor space of a cryogenic dewar at a stable temperature and for preventing temperature excursions that may otherwise occur during refilling of a dewar with liquid cryogen. In some implementations, the devices may include a cryogen space holding liquid cryogen and gas, a separate storage space containing only gas, and a separate path for gas to leave the cryogen space during cryogen refills without passing through or substantially disturbing the temperature of the storage space. Some implementations further provide for passage of gas from the cryogen space to the storage space between cryogen refills to improve cryogen utilization efficiency.

Abstract: Methods and compositions are provided for the cryopreservation of human organs and tissues. In certain embodiments, Step 1 comprises perfusion with a vitrifiable cryoprotectant solution at a temperature above ?10° C. for a time insufficient for the approximate osmotic equilibration of the organ with the solution, followed by cooling the organ to below ?10° C. by perfusion with said solution at a reduced temperature. In certain embodiments, Step 2 comprises increasing the concentration of cryoprotectant further at a temperature from ?10 to ?40° C. In certain embodiments, Step 3 comprises cooling and vitrifying the organ, rewarming it, and perfusing the organ with a vitrifiable concentration of cryoprotectant whose temperature is either raised gradually or is held at ??15° C. Compositions are provided that allow safe organ perfusion with vitrifiable media at >?10° C. and almost complete avoidance of chilling injury at ?20 to ?25° C. and that allow slow warming after vitrification without freezing.

Abstract: A cryogenic storage system for cryogenic storage using liquid refrigerant is provided. The cryogenic storage system includes first and second vacuum vessels, a vacuum source, a quantity of a liquid refrigerant, and at least one temperature control assembly. An insulating wall may be provided on an interior surface of one of the vessels. A common vacuum condition is provided in voids that are present in the wall of the first and second vessels as well as in the insulating wall. The at least one temperature control assembly includes a power supply, a temperature sensor, a heater, and a conductive element. The at least one conductive element provides a link or thermal coupling between a space defined in the first vessel and the liquid refrigerant stored in the second vessel.

Abstract: The present invention provides devices and methods for the cryogenic storage of biological material. Devices of the invention are useful for storing material at a cryogenic temperature. The devices include a temperature chamber defined by a thermally-conductive container and at least one layer of thermal insulation surrounding the thermally-conductive container. Some embodiments utilize one or more heat sources thermally connected to the thermally conductive container. Other embodiments are arranged so that no net flow of heat occurs from the temperature chamber when the temperature chamber is at a set target temperature. Also provided are methods of using the devices.

Abstract: Methods and compositions are provided for the introduction and washout of vitrifiable concentrations of cryoprotectant in organs and tissues. The methods comprise cooling the organ to below ?10° C. by perfusion with a solution having a freezing point below ?10° C., a temperature from ?10 to ?40° C., and a tonicity from 1.1 to 2.0 times isotonic, after previous perfusion with said solution for a time insufficient for approximate osmotic equilibration of the organ with the solution. The methods further comprise increasing the concentration of cryoprotectant further at a temperature from ?10 to ?40° C. to prepare the organ or tissue for vitrification. The methods further comprise cooling and vitrifying the organ, rewarming it, and perfusing the organ with a vitrifiable concentration of cryoprotectant that is the same as or less than the concentration used for vitrification, without the addition of an osmotic buffering agent. Rewarming is accomplished either by rapid (>1°C./min, and preferably ?0.2-20° C.

Abstract: A cryogenic storage system for cryogenic storage using liquid refrigerant is provided. The cryogenic storage system includes first and second vacuum vessels, a vacuum source, a quantity of a liquid refrigerant, and at least one temperature control assembly. An insulating wall may be provided on an interior surface of one of the vessels. A common vacuum condition is provided in voids that are present in the wall of the first and second vessels as well as in the insulating wall. The at least one temperature control assembly includes a power supply, a temperature sensor, a heater, and a conductive element. The at least one conductive element provides a link or thermal coupling between a space defined in the first vessel and the liquid refrigerant stored in the second vessel.

Abstract: A cryogenic storage system for cryogenic storage using liquid refrigerant is provided. The cryogenic storage system includes first and second vacuum vessels, a vacuum source, a quantity of a liquid refrigerant, and at least one temperature control assembly. An insulating wall may be provided on an interior surface of one of the vessels. A common vacuum condition is provided in voids that are present in the wall of the first and second vessels as well as in the insulating wall. The at least one temperature control assembly includes a power supply, a temperature sensor, a heater, and a conductive element. The at least one conductive element provides a link or thermal coupling between a space defined in the first vessel and the liquid refrigerant stored in the second vessel.

Abstract: The present invention provides devices and methods for the cryogenic storage of biological material. Devices of the invention are useful for storing material at a cryogenic temperature. The devices include a temperature chamber defined by a thermally-conductive container and at least one layer of thermal insulation surrounding the thermally-conductive container. Some embodiments utilize one or more heat sources thermally connected to the thermally conductive container. Other embodiments are arranged so that no net flow of heat occurs from the temperature chamber when the temperature chamber is at a set target temperature. Also provided are methods of using the devices.

Abstract: A cryogenic storage system for cryogenic storage using liquid refrigerant is provided. The cryogenic storage system includes first and second vacuum vessels, a vacuum source, a quantity of a liquid refrigerant, and at least one temperature control assembly. An insulating wall may be provided on an interior surface of one of the vessels. A common vacuum condition is provided in voids that are present in the wall of the first and second vessels as well as in the insulating wall. The at least one temperature control assembly includes a power supply, a temperature sensor, a heater, and a conductive element. The at least one conductive element provides a link or thermal coupling between a space defined in the first vessel and the liquid refrigerant stored in the second vessel.

Abstract: Linear polymers of glycerol can prevent or delay ice nucleation in a variety of contexts. Polyglycerol can also be employed in combination with other ice control agents, such as polyvinyl alcohol/polyvinyl acetate copolymers and antifreeze proteins, to provide antinucleation effects that are superior to those of either polyglycerol or the co-antinucleator alone. Polyglycerol has a number of advantageous physical and toxicological properties, such as extreme water solubility, non-toxicity to human beings, non-toxicity to animal tissues and organs in vitro even at extreme concentrations, minimal foaming tendency, minimal retention on hydrophobic surfaces, and stability in solution without the need for periodic heating to reactivate its antinucleation properties.

Abstract: Linear polymers of glycerol can prevent or delay ice nucleation in a variety of contexts. Polyglycerol can also be employed in combination with other ice control agents, such as polyvinyl alcohol/polyvinyl acetate copolymers and antifreeze proteins, to provide antinucleation effects that are superior to those of either polyglycerol or the coantinucleator alone. Polyglycerol has a number of advantageous physical and toxicological properties, such as extreme water solubility, non-toxicity to human beings, non-toxicity to animal tissues and organs in vitro even at extreme concentrations, minimal foaming tendency, minimal retention on hydrophobic surfaces, and stability in solution without the need for periodic heating to reactivate its antinucleation properties.

Abstract: A cryoprotectant solution used for preserving biological material comprising cells is disclosed. The solution comprises dimethyl sulfoxide, an amide such as formamide, urea, acetamide, hydroxyurea, N-methyl formamide, and ethylene glycol or ethylene glycol in combination with propylene glycol wherein the propylene glycol replaces less than 8% w/v of the ethylene glycol.

Abstract: Polyvinyl alcohol and related compounds are provided that inhibit the freezing of water and water solutions. These synthetic compounds preferentially bind and inhibit ice nucleating surfaces in a manner similar to natural antifreeze proteins. The resulting inhibition allows water and water solutions to supercool without ice formation to temperatures below the thermodynamic freezing point. The freezing inhibition occurs at concentrations as small as one part per million, although concentrations up to one part per hundred are preferred. These polyvinyl alcohol additives are very useful for enhancing the performance of antifreeze formulations, biological cryopreservation solutions, and for preventing frost damage to plants and other industrial products and processes.