Capillary assisted vitrification processes and devices
Inventors
MOHANTY, PRAVANSU S. • Chakraborty, Nilay
Assignees
Publication Number
US-12075773-B2
Publication Date
2024-09-03
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
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. A process as provided herein includes providing a membrane comprising a plurality of capillary channels having a first opening and a second opening; providing a vitrification mixture comprising one or more biological materials and a vitrification medium; contacting the membrane with the vitrification mixture such that the first opening of the capillary channels is operably in contact with the vitrification mixture; and desiccating away the vitrification mixture above cryogenic temperature within the plurality of capillary channels.
The invention addresses limitations of prior art vitrification and desiccation methods by providing a method for fast and uniform moisture removal during non-cryogenic vitrification of biological materials and devices to perform the method efficiently. The background describes the need to overcome slow and non-uniform evaporative drying, glassy skin formation, cellular injury during desiccation, and the need for improved vitrification medium and a fast desiccation method with improved cell viability to facilitate long term storage at non-cryogenic temperatures.
Claims Coverage
One independent claim was identified. The main inventive features extract the core elements of the independent process claim.
Membrane comprising a plurality of capillary channels
providing a membrane comprising a plurality of capillary channels, said capillary channels having a first opening and a second opening
Vitrification mixture comprising biological materials and a vitrification medium
providing a vitrification mixture, said vitrification mixture comprising one or more biological materials and a vitrification medium
Operable contact of capillary first openings with vitrification mixture
contacting said membrane with said vitrification mixture such that the membrane contacts a surface of the vitrification mixture, the membrane contacting the vitrification mixture such that the first opening of said capillary channels is operably in contact with said vitrification mixture
Second openings in communication with a lower humidity atmosphere
said second openings of said capillary channels directly operably in communication with a surrounding atmosphere having humidity below that of said vitrification mixture
Desiccation above cryogenic temperature sustaining a liquid network until glassy state
desiccating away the said vitrification mixture above cryogenic temperature within said plurality of capillary channels such that vitrification mixture flow into the plurality of capillary channels sustains a liquid network at an evaporating surface until substantially all of the said vitrification mixture enters into a glassy state
The independent claim covers a capillary-assisted, above-cryogenic desiccation process that combines a capillary-channel membrane, a vitrification mixture containing biological material and vitrification medium, operable contact of the capillary openings with the mixture, exposure of the capillary second openings to a lower-humidity atmosphere, and desiccation within the capillaries that sustains a liquid network until the mixture reaches a glassy state.
Stated Advantages
Fast and uniform moisture removal during non-cryogenic vitrification of biological materials.
Fast and uniform desiccation of a vitrification mixture that can achieve very low and uniform final moisture levels across the sample.
Enables vitrification of larger volumes and enhanced capillary contact area allowing very fast desiccation from multiple sides.
Preservation of cell membrane integrity and improved cell viability at extremely low moisture levels when desiccation is performed by the capillary assisted method.
Enhanced retention of protein activity after desiccation, exemplified by retained insulin activity following the disclosed vitrification method.
Vitrified biological materials can be sealed in protective enclosures impervious to water and air and stored at temperatures between −196° C. to +60° C., including storage at ambient temperatures without requiring a cold chain.
Documented Applications
Preservation and storage of biological materials including proteins, cells, tissues, organs, and cell-based constructs.
Vitrification and storage of mammalian cells and other biological materials such as human mesenchymal stem cells, murine fibroblast cells, blood platelets, bacteria, viruses, mammalian cell membranes, liposomes, enzymes, and combinations thereof.
Vitrification of reproductive cells including sperm cells, spermatocytes, oocytes, ovum, embryos, germinal vesicles, or combinations thereof.
Stabilization of proteinaceous materials and vaccines, with examples including insulin, interleukin 1, interleukin 2, tetanus and hepatitis vaccines.
Sealing vitrified biological samples in protective packages for transportation and long term storage, including storage above cryogenic temperature or subsequent storage at cryogenic temperatures.
Use in wearable device configurations to apply capillary assisted vitrification in situ.
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