Systems and methods for cryopreservation of cells
Inventors
Assignees
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Publication Number
US-10271543-B2
Publication Date
2019-04-30
Expiration Date
Abstract
A fluid sample vessel includes inlet and vent tube fittings formed at one end of a container with an opposite open end closed by a needle septum. A support cap is removably engaged to the container to support the container and protect terminal ends of inlet and vent tubular branches coupled to the fittings. The support cap includes a pair of opposite legs with outwardly directed tabs for mounting within a centrifuge while supporting the cryopreservation container.
Core Innovation
The disclosure provides cryopreservation systems and devices that use an auto-nucleating ice-induction device together with a sterol, including cholesterol, associated with an ice-nucleating composition in a hollow tube. The system uses solution-permeable, cell-impermeable membranes to separate the ice-induction device from cells while allowing controlled ice formation in cryopreservation conditions.
The disclosure reports improved post-thaw semen motility when sterol-bound nucleating devices are used, compared with controls. It further reports no observed cytotoxicity in the testing described, and it discusses reduction of supercooling in relation to ice nucleation effects.
The disclosure also integrates the auto-nucleating ice-induction device into multiple container and sealed-handling formats using hollow interiors, tubular fittings, and outlet openings closed by septa adapted to be pierced by a needle for sterile removal. Flexible heat sealable tubing and sealed pathways are used in container systems, including formats with inlet and vent fittings and an optional gas-permeable sterile micro-filter, to maintain a sealed condition under cryopreservation conditions.
Claims Coverage
The independent claims cover cryopreservation devices and sample container devices that combine sealed container architecture with hollow interior and tubular fittings, heat-sealable flexible tubing forming and maintaining heat seals under cryopreservation conditions, and needle-pierceable septums for sterile removal. Across the independent claims, the main inventive features include device-level structure for sealed handling, passageways, and cryogenic heat-seal survivability, plus container geometry options such as inlet and vent fittings, a graduated storage tube, a second flexible branch, and a gas-permeable sterile micro-filter.
Sealed hollow container with tubular fittings and needle-pierceable septum
A cryopreservation device comprising a container for receiving and storing a liquid sample in a sealed condition under cryopreservation conditions, the container having a hollow interior, a first tubular fitting defining a first passageway communicating with the hollow interior, a second tubular fitting defining a second passageway communicating with the hollow interior, and an outlet opening defining a third passageway communicating with the hollow interior, where the outlet opening is closed by a septum adapted to be pierced by a needle for sterile removal of a stored liquid.
Heat-sealable flexible tubing forming and maintaining a heat seal under cryopreservation conditions
A cryopreservation device wherein a first flexible branch comprising a length of heat sealable tubing connected to the first tubular fitting forms a heat seal and maintains the heat seal under cryopreservation conditions, and further defines the first passageway in communication with the interior of the container.
Cryogenic heat-seal withstand capability
A cryopreservation device wherein the first length of heat sealable flexible tubing is configured to form a heat seal that withstands temperatures as low as -196° C.
Elongated sample container with inlet and vent tube fittings and needle-pierceable septum
A sample container device comprising a container for receiving a sample, the container having an elongated body with a hollow interior from a first end opposite a second end, the first end having an inlet tube fitting and a vent tube fitting opening to the hollow interior, the second end closed by a septum adapted to be pierced by a needle for sterile removal of a stored liquid.
Heat-sealable tubing coupled to inlet fitting for cryogenic heat-seal survivability
A sample container device with a first length of heat sealable tubing coupled to the inlet tube fitting, wherein the heat sealable flexible tubing is configured to form a heat seal that withstands temperatures as low as -196° C.
Graduated storage tube in a sealed cryopreservation container
A cryopreservation device wherein the container comprises a graduated storage tube defining a hollow interior, and wherein the container includes first and second tubular fittings defining first and second passageways communicating with the hollow interior, an outlet opening defining a third passageway communicating with the hollow interior, and a septum closing the outlet opening adapted to be pierced by a needle for sterile removal of a stored liquid.
Second flexible branching passageway with cryogenic heat-seal capability
A cryopreservation device wherein the first flexible branch further includes a second flexible branch comprising a length of heat sealable tubing connected to a second tubular fitting, the second length configured to form a heat seal and maintain the heat seal under cryopreservation conditions, and defining a second passageway in communication with the interior of the container.
Gas-permeable sterile micro-filter in a passageway
A cryopreservation device wherein the second passageway includes a gas-permeable sterile micro-filter disposed therein.
Collectively, the independent claims focus on a sealed cryopreservation container architecture with hollow interior and multiple tubular-defined passageways, flexible heat-sealable tubing that forms and maintains heat seals under cryopreservation conditions, and a needle-pierceable septum for sterile removal. Additional independent-claim scope includes inlet and vent tube fittings in an elongated sample container device, a graduated storage tube, a second heat-sealed flexible branch, and a gas-permeable sterile micro-filter in a passageway.
Stated Advantages
Increases freezing point and reduces supercooling.
Improved post-thaw semen motility.
No observed cytotoxicity in the testing described.
Documented Applications
Cryopreservation of a liquid sample, including post-thaw evaluation in semen motility context.
Sterile removal of a stored liquid from a sealed cryopreservation container using a needle adapted to pierce a septum.
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