Capillary assisted vitrification processes and devices
Inventors
MOHANTY, PRAVANSU S. • Chakraborty, Nilay
Assignees
Publication Number
US-11576374-B2
Publication Date
2023-02-14
Expiration Date
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Abstract
Disclosed are devices and methods for non-cryogenic vitrification of biological materials that include the steps of providing one or more capillary channels of which a first opening is operably in contact with a moisture containing vitrification mixture made of a biological material and a vitrification agent.
Core Innovation
Embodiments of the present invention provide devices and methods for non-cryogenic vitrification of biological materials that include providing one or more capillary channels of which a first opening is operably in contact with a moisture containing vitrification mixture made of a biological material and a vitrification agent. The invention provides a method for fast and uniform moisture removal during non-cryogenic vitrification of biological materials by capillary assisted fast drying. The method desiccates the vitrification mixture by capillary action above cryogenic temperature until the vitrification mixture enters into a glassy state.
The disclosure provides a vitrification composition to preserve the structural integrity (physiological and/or molecular) of the biological material during fast drying, where the vitrification composition comprises trehalose, glycerol and ionic buffer containing one or more large organic ions such as choline and betine. The disclosure provides a non-cryogenic vitrification device including a receptacle comprising a membrane having a plurality of contiguous capillary channels each having a first opening and a second opening, the membrane arranged so as to be able to contact a vitrification mixture such that the first opening of said capillary channels is operably in contact with said vitrification mixture, and an enclosure operably in communication with the second openings of the capillary channels wherein the pressure, temperature and humidity within the enclosure can be controlled. The disclosure further provides a vitrification and long term storage protocol comprising placing biological materials with vitrification medium in a receptacle comprising a plurality of contiguous capillary channels, employing the vitrification method, packaging the receptacle with vitrified biological materials in a protective enclosure and storing the package between −196° C. to 60° C.
Claims Coverage
One independent claim is identified and five main inventive features are extracted from that claim.
Membrane comprising a plurality of contiguous capillary channels
A membrane comprising a plurality of contiguous capillary channels, each of the capillary channels having a first opening and a second opening.
Vitrification mixture comprising biological materials and a vitrification medium
A vitrification mixture that comprises one or more biological materials and a vitrification medium.
Operable contact of membrane with vitrification mixture
Contacting the membrane with the vitrification mixture such that the membrane contacts a surface of the vitrification mixture and the first opening of the capillary channels is operably in contact with the vitrification mixture.
Second openings in direct communication with lower-humidity atmosphere
The second openings of the capillary channels are directly operably in communication with a surrounding atmosphere having humidity below that of the vitrification mixture.
Desiccation by capillary action above cryogenic temperature to a glassy state
Desiccating away the vitrification mixture by capillary action above cryogenic temperature until the vitrification mixture enters into a glassy state.
The independent claim focuses on a capillary-channel membrane in operable contact with a vitrification mixture, exposure of the capillary second openings to a lower-humidity surrounding atmosphere, and desiccation by capillary action above cryogenic temperature until the mixture becomes glassy.
Stated Advantages
Fast and uniform moisture removal during non-cryogenic vitrification of biological materials.
Preservation of structural integrity (physiological and/or molecular) of biological material during fast drying.
Ability to perform non-cryogenic vitrification and to enclose vitrified material for storage at temperatures between −196° C. and +60° C.
Enhanced retention of cell membrane integrity and retention of protein activity (illustrated for insulin) after desiccation using the disclosed vitrification methods and media.
Capability to achieve very fast desiccation rates and ultra-low moisture levels with improved uniformity compared to conventional evaporative drying approaches.
Documented Applications
Preservation and storage of biological materials including proteins, cells, tissues, organs, cell-based constructs, or combinations thereof.
Vitrification of mammalian cells and reproductive cells including sperm cells, spermatocytes, oocytes, ovum, embryos, germinal vesicles, or combinations thereof.
Storage and transport of vitrified biological samples sealed in protective packages and long-term storage at temperatures between −196° C. to +60° C.
Stabilization and desiccation of proteinaceous materials exemplified by insulin and references to interleukin 1, interleukin 2, tetanus and hepatitis vaccines.
Use with a non-cryogenic vitrification device comprising a receptacle with capillary membrane and an enclosure with controllable pressure, temperature and humidity.
Application to a range of biological materials explicitly listed in the specification, including human mesenchymal stem cells, murine fibroblast cells, blood platelets, bacteria, viruses, mammalian cell membranes, liposomes, enzymes, whole blood, red blood cells, white blood cells, platelets, algae, and fungi.
Configuration of the device as a wearable device is described as a possible configuration in the specification.
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