Cryogenic Storage Container

Abstract

Containers for cryogenic storage of biological material are disclosed. Each container is configured to provide a substantially consistent freezing profile throughout the chamber of the container. Specific embodiments include containers having uniform dimensions between two opposing walls defining the chamber, and containers having a ratio of the sum of the interior surface area to the volume of the chamber that is greater than or equal to 1:5 and less than or equal to 2:5.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims one or more inventions disclosed in U.S. Provisional Patent Application No. 63 / 307,614, entitled "Cryogenic Storage Container," filed February 7, 2022. The benefit of this U.S. provisional application under 35 U.S.C. § 119(e) is hereby claimed, and the aforementioned application is incorporated herein by reference. [Background technology]

[0002] The present invention relates broadly to devices for the cryogenic storage of cells and / or tissues, and more specifically to cryogenic storage vessels (i.e., "cryogenic storage vessels") useful for the cryogenic storage of biological material (e.g., cells and / or tissues), such as mammalian cells and tissue samples / specimens.

[0003] Cells and tissues are often cryopreserved to temporarily extend their viability and usefulness in biomedical applications. The process of cryopreservation involves, in part, placing cells in an aqueous solution containing electrolytes and chemical compounds (cryoprotectants) that protect the cells during the freezing process. However, the freezing process is not as harmless as it may be assumed and generally produces extreme changes in chemical, thermal, and electrical properties that can be expected to alter intracellular organelles, cell membranes, and the delicate cell-cell interaction systems associated with tissues and organs.

[0004] When an aqueous solution containing cryoprotectant and cells is cooled to just below its freezing point, the solution remains liquid. This state, where the solution remains liquid below the phase transition temperature, is called supercooling. As the aqueous solution is cooled further below its freezing point, the degree of supercooling increases. Without intervention, the water molecules in the solution will spontaneously crystallize, usually at up to 15°C below the freezing point, and pure water will precipitate as ice.

[0005] During the transition from the liquid to the solid state, the solution passes from a higher to a lower free energy state, resulting in an exothermic reaction. The heat generated during this phase transition causes a temporary heating of the sample, during which the temperature of the sample increases, while the surrounding environment (e.g. the device in which the sample is cryogenically stored) is kept at a constant temperature or is continuously cooled (depending on the cooling method used).

[0006] As the sample heat dissipates, the thermal imbalance between the sample and the cooling device created during this event causes the sample to undergo a rapid cooling rate to re-establish thermal equilibrium. This rapid cooling rate often leads to intracellular ice formation, usually resulting in cell death. This intracellular ice formation usually depends on the mass of the sample, the heat transfer properties of the sample container, the cooling protocol used, and the basic cryobiological properties of the cells.

[0007] In particular, deviations from cooling and / or thawing rates prescribed for a particular cell type may result in cell damage and / or cell death during the process. Conventional cryogenic storage containers useful for directly holding biological material for cryogenic storage have shapes and sizes that allow or result in deviations in cooling and / or thawing rates throughout the volume of the biological material. For example, vials traditionally used for directly holding biological material during cryogenic storage may result in significant variations in cooling and / or thawing rates throughout the volume of the biological material contained therein. Vials often hold multiple samples and are placed within cryogenic storage / freezer boxes that aid in the placement and retrieval of vials from cryogenic storage. Some boxes help to limit the freezing rate of biological samples placed within the vials. For example, some cryogenic storage / freezer boxes are formed of foam materials that provide thermal insulation. However, the freezing profile of the samples within the vials is still affected by the shape and size of the vials.

[0008] In small cylindrical vials (e.g., large enough to hold 5 ml or less), substantially all of the sample freezes at the same rate. However, in larger diameter cylindrical vials (e.g., large enough to hold 10 ml or more, 20 ml or more, 30 ml or more, 40 ml or more, and / or 50 ml or more), it takes substantially longer to freeze the middle portions of these larger diameter vials. Thus, within these larger diameter vials, these middle portions will have different cell viability outcomes than the other portions. Summary of the Invention [Problem to be solved by the invention]

[0009] There remains a need for an apparatus that reduces and / or avoids the problems associated with the above-mentioned unbalanced conditions. Accordingly, there is a need for improvements in this area. [Means for solving the problem]

[0010] The cryogenic storage containers disclosed herein are arranged to provide a substantially consistent freezing profile throughout the chambers of the container, facilitating freezing of the contents of the cryogenic storage container, such as cells, at a constant or more consistent rate, reducing the likelihood of cell damage during freeze and / or thaw cycles (for brevity, the terms "freeze cycle," "freeze profile," or another similar term may be used herein; however, it should be understood that the containers of the present disclosure may provide similar advantages for thaw cycles, thaw profiles, and the like).

[0011] The cryogenic storage containers disclosed herein can have uniform feature dimensions and high dimensional aspect ratios while facilitating or maintaining freeze / thaw consistency and while having relatively large volumes compared to traditional or conventional cryogenic storage vials and bags. For example, the cryogenic storage containers disclosed herein can be configured to hold 30 ml or more.

[0012] Some embodiments of the cryogenic storage container may include a serpentine chamber for holding a volume of liquid, a vent opening and a drain opening each communicating with the serpentine chamber, a filter associated with the vent opening for filtering air entering and exiting the serpentine chamber when communicating with the surrounding environment, and a first end region and a second end region. The serpentine chamber may be at least partially defined by a first wall and a second wall, where the first wall follows a serpentine path from the first end region of the cryogenic storage container to the second end region of the cryogenic storage container. The serpentine path may be defined by a series of straight portions forming a zigzag or chevron pattern. The serpentine path may additionally or alternatively be at least partially defined by a series of bends that bend in alternating directions. The series of bends may be separated by straight portions.

[0013] In some embodiments, the second wall can be spaced apart from the first wall along the serpentine path such that the chamber has a uniform width between the first and second walls along the serpentine path. For example, the width of the chamber between the second and first walls can be between 5 mm and 7 mm, measured along a vector perpendicular to the first wall.

[0014] In some embodiments, the cryogenic storage container may include a heat-sealable tube attached to the drain opening and / or the vent opening, which may hold a filter, especially if the heat-sealable tube is attached to the vent opening.

[0015] In some embodiments, the vent opening can be located at a first end of the serpentine chamber and / or the drain opening can be located at a second end of the serpentine chamber. The serpentine path can have a length extending from the first end to the second end. The length can be greater than the width and / or height of the serpentine chamber.

[0016] In some embodiments, the cryogenic storage container may include a hanger opening positioned to receive an IV bag hanger. The hanger opening may be positioned at an end of the cryogenic storage container opposite the drain opening such that the drain opening is at the bottom of the serpentine chamber when the cryogenic storage container is non-rigidly suspended from the IV hanger. The hanger opening may be off-center along the end of the cryogenic storage container.

[0017] A cryogenic storage container of the present disclosure may comprise a chamber having a fixed volume, the chamber being defined by a wall portion of the cryogenic storage container, the wall portion having an inner surface facing the chamber and an outer surface opposite the inner surface, each inner surface having a surface area, the ratio of the sum of the surface areas to the volume of the chamber being greater than or equal to 1:5 and less than or equal to 2:5. The wall portion defining the chamber may follow a serpentine path from a first end of the chamber to a second end of the chamber. The chamber may have a length extending from the first end to the second end and a uniform width along the length. The width may be greater than or equal to 5 mm and less than or equal to 7 mm. The ratio may be greater than or equal to 3:10 and less than or equal to 2:5. In any of the configurations of the present disclosure, the ratio may be about 1:3.

[0018] The wall portion of the container may have a thermal conductivity of 0.16×10-3 Watts / Kelvin or more and 0.32×10-3 Watts / Kelvin or less at room temperature. In any of the arrangements of the present disclosure, the wall portion may comprise a material having a thermal conductivity of 0.10 W / m·K or more and 0.20 W / m·K or less at room temperature. The body and / or lid may be formed of a polymer, such as a cyclic olefin copolymer. The polymer may be doped with nanoparticles (such as a metal) to increase the thermal conductivity of the material.

[0019] Advantageously, the exemplary cryogenic storage container shown in the figures provided can be oriented in multiple directions. For example, the cryogenic storage container can be positioned with its feet resting on a support surface and the vent and drain openings of the lid located at the top of the cryogenic storage container. The cryogenic storage container may also be oriented such that the drain opening is located at the bottom of the chamber such that substantially all of the liquid contained within the chamber is drained through the drain opening. In such an arrangement, the vent opening is also located at the top of the chamber, allowing air to enter the chamber without mixing with the draining liquid and / or allowing liquid to be expelled from the chamber through the vent opening. During a drain operation, air entering the chamber through the vent opening is filtered by a filter associated with the vent opening to avoid contamination of the interior volume of the serpentine chamber.

[0020] Those skilled in the art will appreciate that the cryogenic storage container may be formed by a variety of manufacturing techniques, such as injection molding, blow molding, rotational molding, gas-assisted molding, and rapid prototyping, to name a few non-limiting examples. The body and lid may be manufactured separately and then attached together, such as by adhesives, ultrasonic welding, hot plate welding, infrared welding, and / or laser welding. In some manufacturing techniques (e.g., rapid prototyping), the lid and body may be manufactured as a single piece. The cryogenic storage container in the illustrated embodiment may be formed by at least injection molding and rapid prototyping. Preferably, regardless of the manufacturing technique, the container is sealable.

[0021] The cryogenic storage containers of the present disclosure preferably have rigid walls that maintain the same shape whether filled or drained, and regardless of the volume of contents stored. Such an arrangement also allows for a more consistent surface area to volume ratio of fluid within the container, as compared to bags or other flexible storage containers. Advantageously, such an arrangement provides greater predictability and reliability during freeze and thaw cycles, helping to promote cell survival.

[0022] Additionally, rigid containers are less prone to leaking than bags and may be preferred in situations where the stored contents are toxic or hazardous. One or more of the interior surfaces of the container walls (e.g., the interior surfaces of the first wall and the second wall) may be hydrophobic. Advantageously, such an arrangement can reduce cell adhesion.

[0023] Nucleation sites for ice crystal formation may be included on the inner surface of the wall defining the chamber. For example, the inner surface may include a coating of ice nucleating agent (INA). Such coating may be protected by a water-permeable top layer (e.g., titanium carbide, molybdenum disulfide with tungsten chalcogenide, and / or boron nitride with graphene). "Seeding" ice formation can reduce and / or eliminate random formation of ice in the liquid being frozen, enhancing sample viability. Random ice formation can occur at unpredictable temperatures. As a result, sample viability can vary greatly between repeated tests using the same freezing protocol. Furthermore, the extremely rapid crystallization that results when ice forms in a highly supercooled solution can be damaging to cells and tissues. Nucleation sites allow control and increase consistency of freezing profiles across samples and batches.

[0024] Advantageously, the arrangements disclosed herein can provide a cryogenic storage vessel having a chamber with a constant ratio of surface area per volume along the length, width, and / or height of the chamber. For example, the chamber can have a volume defined by wall portions of the cryogenic storage vessel, where the wall portions of the cryogenic storage vessel each have an inner surface facing the chamber and an outer surface opposite the inner surface, each inner surface having a surface area, and where the ratio of the sum of the surface areas to the volume of the chamber is greater than or equal to 1:5 and less than or equal to 2:5.

[0025] The containers of the present disclosure may also exhibit greater compressive strength than straight containers having the same wall thickness. Advantageously, the serpentine path of the container's sidewall increases the compressive strength of the structure in at least one direction compared to straight-walled containers.

[0026] The walls of the vessel may include one or more thermocouples. The thermocouple(s) may also be embedded in the walls. In some embodiments, one or more walls of the vessel may include one or more attachment features to facilitate consistent and reliable coupling of the thermocouples to the vessel. The thermocouple(s) may be isolated from the sample so as not to interfere with the nucleation behavior.

[0027] The container may include electrodes arranged to apply high electric fields in the kilovolt range for the purposes of electrofreezing and / or electroporation. Such electrodes may be coupled to one or more walls of the container.

[0028] The vessel may include a displacement body that protrudes into the vessel chamber (e.g., a serpentine chamber). For example, one or more fins or rods may extend into the chamber to displace a volume of the sample therein. Advantageously, such an arrangement may further reduce the distance between a portion of the sample and a surface of the vessel. The displacement body(s) may be arranged to remove heat from the sample. For example, the displacement body(s) may have a thermal conductivity greater than that of the vessel wall. As a specific example, the displacement body(s) may comprise a metal and the vessel wall may comprise a plastic. A thermocouple may also extend into the sample volume defined by the vessel wall.

[0029] The vessel can include an identification tag. Such a tag may include electronics (e.g., RFID, Bluetooth, etc.) for remotely detecting and / or transferring information from the vessel. The vessel can include graduations that indicate the volume of sample in the chamber when the vessel is in a particular orientation.

[0030] The container may be optically transparent or otherwise include structures, properties, and / or materials to facilitate visual inspection of the stored contents, optical quantitative measurements, and other types of non-destructive testing (e.g., spectroscopy, counting, fluorescence, etc.) In some embodiments, the container includes cuvette-like optical properties.

[0031] Methods of using the containers disclosed herein include removing oxygen from the chamber and / or purging the chamber with nitrogen gas through one or more openings; measuring the liquid level electrically, mechanically, optically, and / or acoustically through one or more openings; introducing an INA, a sensor (e.g., optical and / or electrical such as a thermocouple), cryoprotectant, and / or cells into the chamber through one or more openings; placing the container in a cryogenic freezer and / or a cryogenic cell thawing machine such as the VIA Thaw CB1000 from GE Lifesciences.

[0032] The containers disclosed herein can be sealed with the cells and / or cryoprotectant disposed within the chamber. The containers disclosed herein can be provided in a sealed sterile package. [Brief description of the drawings]

[0033] [Figure 1] FIG. 1 is a perspective view of a cryogenic storage vessel according to one embodiment of the present invention. [Diagram 2] FIG. 2 is a perspective view of the body of the cryogenic storage vessel of FIG. 1. [Diagram 3] FIG. 2 is a side view of the cryogenic storage vessel of FIG. 1 in a hanging / draining configuration. [Figure 4] FIG. 2 is a side view of the cryogenic storage vessel of FIG. 1 in a filling configuration. [Diagram 5] FIG. 2 is a side view of the cryogenic storage vessel of FIG. 1 in a drained configuration. [Figure 6] FIG. 2 is a perspective view of the body of the cryogenic storage vessel of FIG. 1. [Figure 7]FIG. 2 is a perspective view of the body of the cryogenic storage vessel of FIG. 1. [Figure 8] FIG. 2 is a perspective view of the body of the cryogenic storage vessel of FIG. 1. [Figure 9] FIG. 2 is a cross-sectional view of the body of the cryogenic storage vessel of FIG. 1. [Figure 10] FIG. 2 is a cross-sectional view of the body of the cryogenic storage vessel of FIG. 1. [Figure 11] FIG. 2 is a perspective view of the lid of the cryogenic storage vessel of FIG. [Figure 12] FIG. 2 is a perspective view of the lid of the cryogenic storage vessel of FIG. [Figure 13] FIG. 13 is a perspective view of a chamber of a cryogenic storage vessel according to another embodiment of the present invention. [Figure 14] FIG. 13 is a perspective view of a chamber of a cryogenic storage vessel according to a further embodiment of the present invention. [Figure 15] FIG. 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 16] FIG. 13 is a perspective view of a chamber of a cryogenic storage vessel according to a further embodiment of the present invention. [Figure 17] FIG. 17 is a partial cross-sectional view of the chamber of FIG. 16. [Figure 18] FIG. 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 19] 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to a further embodiment of the present invention; [Figure 20] FIG. 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 21] 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to a further embodiment of the present invention; [Figure 22] FIG. 22 is a detailed cross-sectional view of the chamber of FIG. 21. [Figure 23] FIG. 13 is a cross-sectional view of a chamber of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 24] FIG. 13 is a perspective view of a chamber of a cryogenic storage vessel according to a further embodiment of the present invention. [Diagram 25]FIG. 13 is a perspective view of a chamber of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 26] FIG. 26 is a partial cross-sectional view of the chamber of FIG. 25. [Figure 27] FIG. 13 is a partial perspective view of a cryogenic storage vessel according to yet another embodiment of the present invention. [Figure 28] FIG. 28 is a schematic partial cross-sectional view of the cryogenic storage vessel and nucleation trigger of FIG. 27. [Figure 29] FIG. 29 is another schematic partial cross-sectional view of the cryogenic storage vessel and nucleation trigger of FIG. 28. [Diagram 30] FIG. 13 is a partial perspective view of a cryogenic storage vessel according to yet another embodiment of the present invention. [Diagram 31] FIG. 31 is a partial perspective view of the cryogenic storage vessel and thermocouple of FIG. [Diagram 32] FIG. 13 is a cross-sectional perspective view of a cryogenic storage vessel according to yet another embodiment of the present invention. [Diagram 33] FIG. 2 illustrates a perspective view of the inside of the bottom wall of a cryogenic storage vessel, according to one embodiment of the present invention. [Diagram 34] FIG. 34 shows a perspective view of the outside of the top wall of the cryogenic storage vessel of FIG. [Diagram 35] FIG. 2 shows a front view of the inside of the bottom wall of a cryogenic storage vessel according to one embodiment of the present invention. [Diagram 36] 36 shows a plan cross-section of the cryogenic storage vessel of FIG. 35. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] For the purpose of promoting an understanding of the principles of the present invention, reference will now be made to the embodiments illustrated in the drawings and specific language will be used to describe these embodiments. It will nevertheless be understood that these are not intended to limit the scope of the invention. Any changes and further modifications in the described embodiments, as well as further applications of the principles of the present invention as described herein, are expected to occur normally to those skilled in the art to which the invention pertains. Although one embodiment of the present invention has been shown in detail, it will be apparent to those skilled in the art that, for the sake of clarity, some features not relevant to the present invention may not be shown.

[0035] The phrases used in the claims and this specification, except for terms expressly defined below, are to have only their plain and ordinary meaning, which plain and ordinary meaning is defined herein to include all consistent dictionary definitions from the most recent published (as of the filing date of this application) commonly used Merriam-Webster dictionary.

[0036] As used herein, the terms "fill", "filling" and "filled" include both partial and complete filling. As used herein, the terms "drain", "draining" and "drained" include both partial and complete draining. As used herein, the term "vent" refers to an opening that allows for equalization of gas pressure between the interior and exterior of the cryogenic storage container. The vent opening may include a flexible membrane (e.g., a bag and / or balloon) that allows for the passage of gas through the vent opening (e.g., through a filter) and / or allows for the expansion and / or contraction of gas within the chamber of the cryogenic storage container. As used herein, the term "chamber width" refers to the distance between the inner surfaces of the two sides of the chamber, as measured along a vector perpendicular (i.e., perpendicular) to at least one of the inner surfaces. The "pipes", "tubes" and "flexible tubes" disclosed herein may comprise materials suitable for cryogenic storage at or below -196°C. Further aspects, objects, features, aspects, benefits, advantages, and embodiments of the present invention will become apparent from the detailed description and drawings provided herein.

[0037] It should be noted that, for the purposes of this specification and the claims, the singular forms "a," "an," "the," etc., include plural referents unless expressly stated otherwise. By way of example, a reference to "a device" or "the device" includes one or more of such devices and equivalents. It should also be noted that directional terms such as "up," "down," "top," "bottom," etc. are used herein solely for the convenience of the reader to aid the reader in understanding the illustrated embodiments, and that the use of these directional terms is not intended in any way to limit the features described, illustrated, and / or claimed to any particular direction and / or orientation.

[0038] The terms "approximately" and "about" when modifying a quantity, size, or shape, unless otherwise specified, are intended to mean any value with a tolerance of ±10% of that value.

[0039] All ranges disclosed herein include all subranges contained therein unless otherwise indicated. For example, a range "less than 10" can include all subranges between a minimum value of 0 and a maximum value of 10 (inclusive), i.e., all subranges with a minimum value of 0 or more and a maximum value of 10 or less, such as 1 to 5. In some cases, the numerical values ​​listed for a parameter may be negative. In this case, exemplary values ​​for the range listed as "less than 10" may be negative, such as -1, -2, -3, -10, -20, -30, etc.

[0040] FIG. 1 illustrates a cryogenic-storage vessel 100 including a body 102. FIG. 2 illustrates a perspective view of the body 102. FIGS. 6, 7, and 8 illustrate perspective views of the body 102 of the cryogenic-storage vessel 100. FIGS. 9 and 10 illustrate cross-sectional views of the body 102 of the cryogenic-storage vessel 100. With reference to FIGS. 1, 2, and 6-10, the body 102 defines a chamber 110 for holding a volume of liquid. The chamber 110 is defined by one or more walls of the body 102. For example, the body 102 can have a first wall 120 extending along a first side 122 of the chamber and a second wall 124 extending along a second side 126 of the chamber 110 opposite the first side 122. The chamber 110 may also be defined by a first end wall 130 disposed at a first end 132 of the chamber 110 and a second end wall 134 disposed at a second end 136 of the chamber 110. Extending along and closing a bottom 140 of the chamber 110 is a bottom wall 142. The walls may be rigid to facilitate shape retention.

[0041] The first wall 120, the second wall 124, and the chamber 110 follow a serpentine path along the length L of the body 102. Each of the first wall 120 and the second wall 124 includes a plurality of wall portions disposed at various angles relative to one another, or at alternating angles from the first end 132 to the second end 136. Along the serpentine path, the first wall 120 and the second wall 124 have a uniform characteristic dimension therebetween, i.e., a uniform distance 156 therebetween, measured along a direction perpendicular to the inner surface 150 of the first wall 120. The uniform distance 156 is uniform throughout a planar cross-section parallel to the distance D. For example, the distance 156 between the second wall 124 and the first wall 120, measured along a vector perpendicular to the inner surface 150 of the first wall 120, may be greater than or equal to 5 mm and less than or equal to 7 mm. The first wall 120 may be parallel to the second wall 124 along a serpentine path.

[0042] The first wall 120 and the second wall 124 may also have a uniform distance 156 therebetween along the height of the body. Thus, the first wall 120 and the second wall 124 may be spaced apart by a uniform distance 156 across the length L and / or height H. Advantageously, having a uniform characteristic dimension between the first wall 120 and the second wall 124 along the length L and / or height H of the body 102, with the uniform characteristic dimension being relatively small in value, helps maintain a substantially uniform freezing profile along the length L and / or height H of the body 102.

[0043] In practice, precise uniformity of uniform feature dimensions is difficult to manufacture, so some tolerance is allowed. Furthermore, in some embodiments, dimples, recesses, protrusions, or other non-uniform structural features may be useful or necessary, for example, to facilitate manufacturing. These non-uniform structural features may affect the uniform feature dimensions across a portion of the wall of the chamber 110. Thus, in some embodiments, the uniform feature dimensions may be within, for example, 0.01 mm, 0.1 mm, 0.5 mm, 1.0 mm, or 2.0 mm. In some embodiments, the uniform feature dimensions may be within, for example, 0.1%, 0.5%, 1%, 2%, 5%, 10%, 20%, or 25% of a uniform feature dimension target or median value. In general, high uniformity of the uniform feature dimensions is desirable to enhance uniform cooling and / or melting rates of the contained biological sample / material.

[0044] In some embodiments, the uniform characteristic dimension may be uniform within any of the tolerances mentioned above for greater than or equal to 90%, greater than or equal to 95%, or greater than or equal to 99% of the vertical distance between the first wall portion 120 and the second wall portion 124.

[0045] In some embodiments, the walls may deviate slightly from one another along a direction extending from the bottom 140 of the body 102 to the top 160 of the body 102. While this deviation may not have a significant effect on changing the freezing profile along the height H of the body 102, it may make it easier for the body 102 to release from a mold during manufacturing.

[0046] Advantageously, a serpentine chamber having a width, height, and overall length between its interior walls can provide a larger liquid storage volume than a linear chamber of the same width, height, and overall length. Thus, a serpentine chamber configured for a cryogenic storage box can store more liquid than a linear container of similar dimensions (i.e., equal overall length).

[0047] The thermal resistance of the first wall 120 and the second wall 124 is preferably uniform along the length L and height H. For example, the first wall 120 and / or the second wall 124 can have a uniform material thickness along the length L and height H. The first wall 120 and the second wall 124 can have the same thermal resistance to promote uniform heat transfer during freeze / thaw cycles.

[0048] 3, the cryogenic storage vessel includes one or more openings positioned to provide access to a portion of the chamber 110. For example, a first opening 202 of the cryogenic storage vessel can be in communication with the first end 132 of the chamber 110, a second opening 204 can be in communication with the second end 136 of the chamber 110, and a third opening 206 can be in communication with either the first end 132 or the second end 136 of the chamber 110.

[0049] 1-3, one or more of the openings 202, 204, 206 may be associated with a connector 207 (e.g., a luer or barb fitting) arranged for coupling to a liquid filling device 212 (e.g., a tube, a syringe, a supply bag, or a pump). For example, in the illustrated embodiment, the connector 207 is a barb fitting configured to mate with resistance and / or interference within a flexible tube. The liquid filling device 212 is shown in FIG. 3 as a flexible tube connected to the barb fitting of the opening 202. The flexible tube may be weldable (e.g., heat sealable) such that the inner surfaces of the flexible tube may be welded together to close the lumen of the flexible tube and seal the corresponding opening in the cryogenic storage vessel 100.

[0050] One or more of the openings 202, 204, 206 may be positioned for filling and / or draining liquid from the chamber 110. In some embodiments, one or more of the openings 202, 204, 206 may be a high-throughput fill port. In some embodiments, at least one of the openings 202, 204, 206 in communication with the chamber 110 may be positioned to supply air to and / or exhaust air from the chamber 110. For example, a vent opening positioned to vent air when liquid is being drained from the chamber 110 allows air to enter the chamber 110, and a vent opening positioned to allow air to exit the chamber 110 when liquid is being filled into the chamber 110. In the illustrated embodiment, a first opening 202 at a first end of the chamber 110 is positioned for venting, a second opening 204 at a second end of the chamber is positioned for draining, and a third opening 206 is positioned for filling the chamber with liquid.

[0051] In some embodiments, the cryogenic-storage vessel 100 can include a lid. Figures 11 and 12 are perspective views of a lid 300 of the cryogenic-storage vessel 100. With reference to Figures 1, 2, 11, and 12, the lid 300 can seal an end or side of the chamber 110. For example, in the illustrated embodiment, the top of the chamber 110 defined by the top 160 of the body 102 is sealed by the lid 300. The lid 300 can include one or more of the openings 202, 204, 206.

[0052] The bottom surface 302 of the lid 300 can include a wall 310 extending between a vent opening (e.g., first opening 202) and a fill opening (e.g., third opening 206). The wall 310 prevents fluid entering through the fill opening from quickly escaping through the vent opening, particularly when a negative pressure is applied to the vent opening to evacuate gas from the chamber 110.

[0053] With reference to FIG. 3, the filter element 220 may be associated with a vent opening and may be arranged to filter air entering the chamber 110 during draining of the chamber 110. The filter element 220 may be a microfilter, more specifically a filter that provides a microbial barrier. Such a microfilter may be, for example, a 3 μm, 0.2 μm, or 0.1 μm sterile porosity microfilter. In other embodiments, the microfilter may have a different structure. The filter element 220 may be arranged to be gas permeable, but it is generally hydrophobic (i.e., resistant to the passage of the liquid sample or cell suspension stored in the container). Thus, the filter element 220 is capable of retaining the suspension within the cryogenic-storage container 100, evacuating gas from the cryogenic-storage container 100 when the cryogenic-storage container 100 is being filled (either partially or completely), and supplying gas to the cryogenic-storage container 100 when the suspension is being withdrawn from the cryogenic-storage container 100. Although the filter element 220 is shown as being positioned / held within the flexible tube 212 associated with the first opening 202, the filter element 220 may be positioned differently in the path of the air through the vent opening.

[0054] The cryogenic storage vessel may include a hanging element 230, which may be a loop as shown in FIGS. 1-3, or another currently known or later developed arrangement for suspending the cryogenic storage vessel above a surface. The hanging element 230 may be positioned such that when the cryogenic storage vessel 100 is suspended from the hanging element 230 as shown in FIG. 3, the drain opening (e.g., opening 204) is located at the lowest point of the chamber 110. Thus, fluid will flow under gravity toward the drain opening. Such an arrangement may facilitate maximum or complete drainage of fluid from the chamber 110, except perhaps some residual fluid such as moisture or fluid film adhering to the interior surface of the chamber 110. For example, the hanging element may be located at an end opposite the drain opening and / or near the side opposite from the side having the drain opening. The hanging element 230 may be off-center along an end of the cryogenic storage vessel 100.

[0055] The cryogenic-storage vessel 100 can be positioned to support itself on a horizontal surface in various positions. For example, the cryogenic-storage vessel 100 can be positioned to support itself in a filled configuration. FIG. 4 is a side view of the cryogenic-storage vessel 100 in a filled configuration. In the filled configuration, the vent opening is higher than at least 80% of the chamber. Thus, during filling, liquid rises toward the vent opening and gas in the chamber is discharged through the vent opening. The cryogenic-storage vessel 100 can also be positioned to support itself in a drained configuration (see FIGS. 3 and 5), which may be the same as or different from the filled configuration. In the drained configuration, the drain opening is lower than at least 80% of the chamber.

[0056] To support itself in one or more configurations (e.g., a fill and / or drain configuration), the cryogenic-storage vessel 100 can include one or more stands. For example, with reference to Figures 2, 4, and 5, the illustrated embodiment of the cryogenic-storage vessel 110 has a first stand 240 at the first end 132 and a second stand 242 at the second end 136. In the fill configuration shown in Figure 4, both the first stand 240 and the second stand 242 contact the support surface 244, and the vent openings and the drain / fill openings are located above the chamber 110. In the drain configuration shown in Figure 5, the cryogenic-storage vessel 100 is supported on the support surface 244 by the second stand 242, and the drain openings are located below the chamber. The inner surface of the bottom wall(s) defining the chamber 110 (e.g., second end wall 134 shown in Figures 5 and 8) slopes downwardly toward a drain opening in a drain configuration and / or the cryogenic storage vessel 100 is supported by a hanging element 230 so that substantially all of the fluid, except perhaps residual fluid (e.g., less than 1% by volume), can be removed from the chamber 110.

[0057] In some embodiments, the cryogenic storage vessel 100 can have a height of about 80 mm, a length of about 125 mm, a width of about 20 mm, and a volume of about 48 ml. In some embodiments, the cryogenic storage vessel 100 has a volume in the range of 100 ml to 250 ml.

[0058] As briefly described above, the relatively large volume cryogenic storage containers of the present invention have uniform feature dimensions and high dimensional aspect ratios to facilitate consistency and uniformity of freeze / thaw rates. Such containers may have serpentine chambers, such as the cryogenic storage container 100 described above. Alternatively, such containers may have other shapes. Some exemplary shapes of chambers for such containers are shown in Figures 13-26 and 32-34.

[0059] 13, there is illustrated a cryogenic storage chamber 400 having a three-dimensional rectangular shape. The chamber 400 has a uniform characteristic dimension R, more specifically a thickness dimension. The chamber 400 also has a height dimension H and a width dimension W. The aspect ratio of the chamber 400 is defined as follows:

number

[0060] The surface area of ​​the chamber 400 is related to the aspect ratio as follows:

number

[0061] For relatively high aspect ratios, or relatively high SA1 to SA2 or SA1 to SA3 ratios, the characteristic dimension R significantly impacts and provides substantial consistency in the freezing profile of the chamber 400. Such relatively high aspect ratios can be, for example, 1:1 to 2:1, preferably 2:1 to 5:1, and more preferably 5:1 to 20:1.

[0062] 14, a cryogenic storage chamber 500 is shown having a cylindrical shape. The chamber 500 has a uniform characteristic dimension R, more specifically, a diameter. The chamber 500 also has a height dimension H. The aspect ratio of the chamber 500 is defined as follows:

number

[0063] The surface area of ​​the chamber 500 is related to the aspect ratio as follows:

number

[0064] For relatively high aspect ratios, or relatively high SA1 to SA2 ratios, the characteristic dimension R significantly impacts and provides substantial consistency in the freeze profile of the chamber 500. Such relatively high aspect ratios can be, for example, 1:1 to 2:1, preferably 2:1 to 5:1, and more preferably 5:1 to 20:1.

[0065] 15, there is shown a cryogenic storage chamber 600 having a three-dimensional hollow cylindrical or donut shape. The chamber 600 has a uniform characteristic dimension R, more specifically the difference between an outer radius TA and an inner radius ra.

[0066] 16 and 17, a cryogenic storage chamber 700 is illustrated having a plurality of connected three-dimensional square X shapes 702. With particular reference to FIG. 17, the chamber 700 has a uniform characteristic dimension R, more specifically the vertical width of each leg of the X shapes 702. The chamber 700 also includes an overall thickness T, an overall width W, and a number n of X shapes 702. Since the X shapes 702 are square, W / n=T. The legs of the X shapes 702 have pointed ends 704. In some embodiments, the optimal value of R / T provides the chamber 700 with a relatively large perimeter without excessively reducing the area of ​​the chamber 700. In some embodiments, the value of R / T is within the range of 0.4-0.5, more specifically about 0.45. In some embodiments, the chamber 700 has a height of about 80 mm, an overall width W of about 120 mm, an overall thickness T of about 20 mm, and a volume of about 103 ml.

[0067] 18, there is illustrated a cryogenic storage chamber 800 having a plurality of connected three-dimensional square X-shapes 802. Chamber 800 is similar to chamber 700 described above, except that the legs of the X-shapes 802 have rounded ends 804.

[0068] 19, a cryogenic storage chamber 900 is illustrated having a plurality of three-dimensional rectangular shapes 902. Each shape 902 includes a uniform characteristic dimension R, more specifically a thickness dimension. The shapes 902 are connected by a plurality of connectors 904.

[0069] 20, a cryogenic storage chamber 1000 is illustrated having a three-dimensional rectangular shape with a plurality of cylindrical voids 1002. The chamber 1000 includes an overall thickness T, an overall width W, and a number of segments n. The chamber 1000 has a uniform characteristic dimension (not labeled), more specifically the vertical width between the cylindrical voids 1002.

[0070] 21 and 22, a cryogenic storage chamber 1100 is illustrated having a plurality of joined three-dimensional hollow hexagonal shapes 1102. With particular reference to FIG. 22, the chamber 1100 has a uniform characteristic dimension R, more specifically the vertical width of each of the hollow hexagonal shapes 1102. The chamber 1100 also includes an overall thickness T, an overall width W, and a number n of the hollow hexagonal shapes 1102.

[0071] 23, there is illustrated a cryogenic storage chamber 1200 having a plurality of three-dimensional hollow cylinders 1202. The chamber 1200 includes an overall thickness T, an overall width W, and a number n of hollow cylinders 1202.

[0072] 24, a cryogenic storage chamber 1300 is illustrated having nested hollow hexagonal shapes. More specifically, the chamber 1300 includes an outer hollow hexagonal shape 1302, an inner hollow hexagonal shape 1304, and a connector 1306 joining the hollow hexagonal shapes 1302, 1304. The chamber 1200 can have a longitudinal height of about 100 mm, a width of about 60 mm, a thickness of about 51 mm, and a volume of about 108 ml.

[0073] 25 and 26, a cryogenic storage chamber 1400 is illustrated having a plurality of cylinders 1402. With particular reference to FIG. 26, each cylinder 1402 includes a uniform characteristic dimension R, more specifically a radius. The cylinders 1402 are connected by a plurality of connectors 1404.

[0074] 27-29, a cryogenic storage vessel 1500 is illustrated, which may have generally the same structure and dimensions as any of the cryogenic storage vessels described herein. Additionally, the cryogenic storage vessel 1500 includes one or more nucleation sites 1502 (illustratively, two nucleation sites) that facilitate ice crystal formation. Illustratively, each nucleation site 1502 is formed as a depression or recess 1504 in a wall 1506 of the cryogenic storage vessel 1500. In some embodiments, the method of freezing a sample facilitated by the nucleation sites 1502 is as follows: First, the sample 1508 is sent to the cryogenic storage vessel 1500. The sample 1508 is then cooled (e.g., by placing in a programmable freezer) to freezing or below freezing (i.e., a supercooled state where water remains liquid but is very close to freezing due to random ice nucleation). Next, as illustrated in Figure 28, a nucleation trigger 1510 (e.g., a metal body cooled to below freezing) is moved toward one of the nucleation sites 1502. As illustrated in Figure 29, the nucleation trigger 1510 then contacts the nucleation site 1502. This causes the nucleation trigger 1510 to initiate ice nucleation at the nucleation site 1502, which then propagates outward from the nucleation site 1502.

[0075] 30 and 31, a cryogenic storage vessel 1600 is illustrated, which may generally have the same structure and dimensions as any of the cryogenic storage vessels described herein. Additionally, one or more walls 1602 of the vessel 1600 may include one or more attachment mechanisms 1604, each of which receives a thermocouple 1606 (FIG. 31). The attachment mechanisms 1604 inhibit the thermocouples 1606 from becoming dislodged from the vessel 1600, which may result in temperature deviations on the manufacturing record. Additionally, the attachment mechanisms 1604 facilitate consistent location of temperature measurements between samples and batches. Illustratively, the attachment mechanisms 1604 are formed as exterior recesses or channels 1608 in the walls 1602 and wire retaining elements 1610.

[0076] Referring to FIG. 32, a cryogenic storage container 1700 is illustrated, which may generally have the same structure and dimensions as any of the cryogenic storage containers described herein. The container 1700 includes opposing upper and lower walls 1702 and 1704 that define a chamber 1705. The uniform characteristic dimension D1 may vary to include one or more recesses for specific purposes other than facilitating consistent and uniform freeze / thaw rates, in this example, as described above. In this example, the cryogenic storage container 1700 facilitates visual inspection, optical quantitative measurements, and other types of non-destructive testing of the contents of the container 1700. The opposing upper and lower walls 1702 and 1704 may be transparent or may include transparent portions that facilitate microscopic inspection and spectroscopic analysis. Such processes in turn facilitate in-process measurements, diagnosis, and early identification of problems with the sample. In some embodiments, the upper wall 1702 and / or the lower wall 1704 include one or more recesses to inhibit surface damage to the respective walls 1702, 1704 during manufacturing, distribution, and / or use.

[0077] Illustratively, the top wall 1702 includes one recess 1706 and the bottom wall 1704 includes two recesses 1708. In some embodiments, the top wall 1702 and the bottom wall 1704 define an optical path length 1710 therebetween, which may be uniform across a series of containers of various volumes. The one recess 1706 and the two recesses 1708 may be deep enough to serve the intended purpose of reducing or preventing damage to the walls 1702, 1704, while also being shallow enough to maintain sufficient uniformity of the uniform feature dimension D1 (e.g., within the tolerances discussed above) and sufficient consistency / uniformity of the freeze / thaw rates within the chamber.

[0078] FIG. 33 shows the outside of the top wall 1802 of the cryogenic storage vessel 1800, and FIG. 34 shows the inside of the bottom wall 1804 of the cryogenic storage vessel 1800. FIGS. 33 and 34 also show the vent line 1820, the fill line 1822, and the spike port 1824. As with the embodiment of FIG. 32, the uniform feature dimension (not shown, but corresponding to D1 in FIG. 33) may vary at specific locations or regions between the top wall 1802 and the bottom wall 1804, in this example by including one or more recesses for a specific purpose other than to promote consistent and uniform freeze / thaw rates. The top wall 1802 includes a first recess 1806, and the bottom wall 1804 includes a second recess 1808 and a third recess 1810. The first recess 1806, the second recess 1808, and the third recess 1810 may be deep enough to serve their intended purpose while also being shallow enough to maintain sufficient uniformity of uniform feature dimensions (e.g., within the tolerances discussed above) and sufficient consistency / uniformity of the freeze / thaw rate within the chamber. For example, the uniform feature dimension, i.e., vertical distance, between the top wall 1802 and the bottom wall 1804 may be a maximum of 10 mm if there are no recesses and a minimum of 8 mm if there are recesses in each wall 1802, 1804. Thus, if the vertical distance between the top wall 1802 and the bottom wall 1804 extends between the two recesses, the uniform feature dimension may be considered to be 10 mm, with a deviation of minus 20%. In this example, the maximum height H of the cryogenic storage vessel 1800 is about 100 mm, resulting in an aspect ratio of about 10 to 1.

[0079] Figure 35 shows the inside of the bottom wall 1904 of the cryogenic-storage vessel 1900, and Figure 36 shows a planar cross section of the cryogenic-storage vessel 1900 parallel to the uniform characteristic dimension D2. The cryogenic-storage vessel 1900 is similar to the cryogenic-storage chamber 1800, except that the top wall 1902 has two recesses, and the recesses are shaped differently.

[0080] While the invention has been illustrated and described in detail in the drawings and foregoing description, the same is to be considered as illustrative in nature and not restrictive, with the understanding that only the preferred embodiment has been shown and described, and that protection is desired for all changes, equivalents, and modifications that come within the spirit of the invention as defined by the following claims. All publications, patents, and patent applications cited in this specification are hereby incorporated by reference as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

Claims

1. 1. A cryogenic storage vessel comprising: A first body defining a chamber, the first body including a plurality of walls, the plurality of walls comprising: a first wall extending along a first side of the chamber; and a first body including a second wall extending along a second side of the chamber, the second side being opposite the first side with respect to the chamber; 1. A cryogenic storage vessel comprising: a chamber having a volume in a range of 25 mL to 250 mL; the chamber having a uniform characteristic dimension extending from the first wall to the second wall perpendicular to the first wall and the second wall, the uniform characteristic dimension being uniform across a planar cross-section of the chamber parallel to the uniform characteristic dimension.

2. 2. The cryogenic storage vessel of claim 1, wherein the uniform characteristic dimension is at least 90% of all distances extending from the first wall to the second wall and perpendicular to the first wall and the second wall.

3. a vent opening in fluid communication with the chamber; and The cryogenic storage vessel of claim 1 further comprising a drain opening in fluid communication with the chamber.

4. 4. The cryogenic storage vessel of claim 3, further comprising a filter associated with said vent opening for filtering air entering and exiting said chamber.

5. 4. The cryogenic storage vessel of claim 3, further comprising a heat sealable tube attached to at least one from the group consisting of said drain opening and said vent opening.

6. The cryogenic storage vessel of claim 3 , wherein the vent opening is located at a first end of the chamber and the drain opening is located at a second end of the chamber.

7. The cryogenic storage vessel of claim 1 , wherein the chamber follows a serpentine path.

8. The body comprises: a first end wall at a first end of the chamber, the first end wall connecting the first wall and the second wall; and a second end wall at a second end of the chamber, the second end wall being on an opposite side of the chamber from the first end wall; The cryogenic storage vessel of claim 7 , wherein the serpentine path extends along a length between the first end wall and the second end wall.

9. 8. The cryogenic storage vessel of claim 7, wherein the first wall includes a plurality of wall portions positioned at different angles relative to one another and the second wall includes a plurality of wall portions positioned at different angles relative to one another, each wall portion of the first wall being substantially parallel to and opposite one of the wall portions of the second wall.

10. 8. The cryogenic storage vessel of claim 7, wherein the serpentine path is defined at least in part by a series of turns that bend in alternating directions, the series of turns being separated by straight wall portions.

11. The cryogenic storage vessel of claim 7 , wherein the serpentine path is defined by a series of straight wall portions arranged in a chevron pattern.

12. 10. The cryogenic storage vessel of claim 1, wherein the uniform characteristic dimension is greater than or equal to 5 mm and less than or equal to 10 mm.

13. The cryogenic storage vessel of claim 1 , further comprising a hanger opening positioned to receive an IV bag hanger.

14. 14. The cryogenic storage container of claim 13, wherein the hanger opening is positioned at an end of the cryogenic storage container opposite the drain opening such that the drain opening is at the bottom of the serpentine chamber when the container is freely hanging from the IV bag hanger.

15. The cryogenic storage vessel of claim 14 , wherein the hanger opening is off-center along an end of the cryogenic storage vessel.

16. 10. The cryogenic storage vessel of claim 1, further comprising nucleation sites to facilitate ice crystal formation.

17. 17. The cryogenic storage vessel of claim 16, wherein the nucleation sites comprise depressions formed in the plurality of walls.

18. The cryogenic storage vessel of claim 1 , wherein the plurality of walls include a mounting mechanism, the cryogenic storage vessel further comprising a thermocouple carried by the mounting mechanism.

19. The cryogenic storage vessel of claim 18 , wherein the attachment feature comprises a recess.

20. 20. The cryogenic storage vessel of claim 19, wherein the attachment mechanism further comprises a wire retaining element.

21. The cryogenic storage vessel of claim 1 , wherein the plurality of walls includes a transparent wall portion.

22. 22. The cryogenic storage vessel of claim 21, wherein the transparent wall portion is concave.

23. A second body defining a second chamber, the second body including a second plurality of walls, the second plurality of walls comprising: a third wall extending along a first side of the second chamber; and a second body including a fourth wall extending along a second side of the second chamber, the second side of the second chamber being on an opposite side of the second chamber from the first side of the second chamber; the second chamber has a second volume in a range of 25 mL to 250 mL, the second chamber has a second uniform characteristic dimension extending from the third wall to the fourth wall perpendicular to the third wall and the fourth wall, the second uniform characteristic dimension being uniform across a planar cross-section of the second chamber parallel to the second uniform characteristic dimension; The cryogenic storage vessel of claim 1 , wherein the first chamber is in fluid communication with the second chamber.

24. A third body defining a third chamber, the third body including a third plurality of walls, the third plurality of walls comprising: a fifth wall extending along a first side of the third chamber; and a third body including a sixth wall extending along a second side of the third chamber, the second side of the third chamber being on an opposite side of the third chamber from the first side of the third chamber; the third chamber has a third volume, the third chamber has a third uniform characteristic dimension extending from the fifth wall to the sixth wall perpendicular to the fifth wall and the sixth wall, the third uniform characteristic dimension being uniform across a planar cross-section of the third chamber parallel to the third uniform characteristic dimension; 24. The cryogenic storage vessel of claim 23, wherein the second chamber is in fluid communication with the third chamber.

25. 1. A cryogenic storage vessel comprising: a chamber having a fixed volume; and a wall portion defining the chamber, the wall portion having an inner surface facing the chamber and an outer surface opposite the inner surface; A cryogenic storage vessel comprising a wall portion, each inner surface having a surface area, the ratio of the sum of the surface areas of the inner surfaces to the volume of the chamber being greater than or equal to 1:5 and less than or equal to 2:

5.

26. 26. The cryogenic storage vessel of claim 25, wherein the wall portions defining the chamber follow a serpentine path from a first end of the chamber to a second end of the chamber.

27. 27. The cryogenic storage vessel of claim 26, wherein the chamber has a length extending from the first end to the second end and a uniform width along the length.

28. 28. The cryogenic storage vessel of claim 27, wherein the width is greater than or equal to 5 mm and less than or equal to 10 mm.

29. The wall portion has a thickness of 0.16×10 －３ Watts / Kelvin or higher 0.32 x 10 －３ 26. The cryogenic storage vessel of claim 25 having a thermal conductivity of less than or equal to Watt per Kelvin.

30. 26. The cryogenic storage vessel of claim 25, wherein the wall portion comprises a material having a thermal conductivity of 0.10 W / m·K or greater and 0.20 W / m·K or less at room temperature.

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