Cryogenic storage container

Abstract

A container for cryogenic storage of biological material is disclosed. Each container is configured to provide a substantially uniform freeze distribution throughout the chamber of the container. Particular embodiments include a container having a uniform size between two opposing walls defining the chamber, and a container wherein the ratio of the sum of internal surface areas to the volume of the chamber is between 1: 5 inclusive and 2: 5 inclusive.

Description

[0001] Citation of Related Applications

[0002] This application claims one or more inventions disclosed in U.S. Provisional Patent Application No. 63 / 307,614, filed on February 7, 2022, and entitled "Cryogenic Storage Container." The benefits of 35 U.S.C. §119(e) under this U.S. Provisional Application are hereby claimed, and the foregoing application is incorporated herein by reference. Background Art

[0003] The present invention generally relates to devices for cryogenic storage of cells and / or tissues, and more particularly to cryogenic storage containers (i.e., "cryostorage containers") useful for cryopreservation of biological materials (e.g., cells and / or tissues) such as mammalian cells and tissue samples / specimens.

[0004] Cells and tissues are often cryopreserved to temporarily prolong 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 gentle as one might think; it generally causes extreme changes in chemical, thermal, and electrical properties that are expected to alter the delicate intercellular interaction systems associated with tissues and organs and the cell organelles within the cell.

[0005] When an aqueous solution containing a cryoprotectant and cells is cooled to a temperature slightly below its freezing point, the solution remains liquid. This situation where such a solution remains liquid below its phase transition temperature is called supercooling. When the aqueous solution is further cooled below its freezing point, the degree of supercooling increases. In the absence of intervention, water molecules in the solution often spontaneously crystallize at no more than 15°C below the freezing point, and pure water will precipitate as ice.

[0006] During the transition from liquid to solid, the solution moves from a higher free energy state to a lower free energy state, resulting in an exothermic reaction. The heat generated during this phase change causes a brief warming of the sample, during which the sample temperature increases. At the same time, the surrounding environment (e.g., a device for storing the sample at low temperature) remains at a constant temperature or continues to cool (depending on the cooling method used).

[0007] Subsequently, as the heat in the sample 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 reestablish thermal equilibrium. In many cases, this rapid cooling rate causes the formation of intracellular ice, which often leads to cell death. The formation of such intracellular ice generally depends on the quality of the sample, the heat transfer properties of the sample container, the cooling scheme used, and the basic cryobiological properties of the cells.

[0008] In some embodiments, the present invention relates to a cryogenic storage container. The cryogenic storage container is a container that is used to store the biomaterial directly at low temperature and / or to store the biomaterial at low temperature. The ...

[0009] In small cylindrical bottles (e.g., large enough to hold 5 ml or less), substantially all samples freeze at the same rate. However, in larger diameter cylindrical bottles (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), the middle portions of the larger diameter bottles freeze substantially longer. Thus, the middle portions have different cell viability results than other portions within the larger diameter bottles.

[0010] There remains a need for means to reduce and / or avoid the problems associated with the unbalanced conditions described above.Therefore, improvements are needed in this area. Summary of the invention

[0011] Containers for cryogenic storage as disclosed herein are arranged to provide a substantially uniform freezing profile throughout the chamber of the container. Such an arrangement facilitates freezing the contents of the cryogenic storage container (such as cells) at a constant rate or a more constant rate to reduce the possibility of damaging the cells during the freezing and / or thawing cycle. (In some examples herein, for the sake of brevity, the term "freezing cycle", "freezing profile" or another similar term is used. However, it should be understood that containers according to the present disclosure can provide advantages similar to thawing cycles, thawing profiles, etc.).

[0012] The cryogenic storage containers disclosed herein can have uniform feature dimensions and high aspect ratios while promoting or maintaining freeze / thaw consistency and at the same time have a relatively large capacity compared to conventional or previously existing bottles and bags for cryogenic storage. For example, the cryogenic storage containers disclosed herein can be arranged to hold 30 ml or more.

[0013] Some embodiments of cryogenic storage containers may include: a serpentine chamber for accommodating a volume of liquid; a vent opening and a discharge opening, each of which is in communication with the serpentine chamber; a filter, which is associated with the vent opening when communicating with the surrounding environment to filter air entering and / or leaving the serpentine chamber; 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, wherein 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 line portions forming a zigzag pattern or a herringbone pattern. The serpentine path may additionally or alternatively be at least partially defined by a series of curves that bend in alternating directions. The series of curves may be separated by straight line portions.

[0014] In some embodiments, the second wall can be spaced apart from the first wall along a serpentine path so that the chamber has a uniform width between the first wall and the second wall along the serpentine path. For example, the width of the chamber between the second wall and the first wall measured along a vector perpendicular to the first wall can be between 5 mm (inclusive) and 7 mm (inclusive).

[0015] In some embodiments, the cryogenic storage container may include a heat-sealed tube attached to the discharge opening and / or the vent opening. The heat-sealed tube may retain a filter, particularly if the heat-sealed tube is attached to the vent opening.

[0016] In some embodiments, the vent opening may be positioned at a first end of the serpentine chamber and / or the exhaust opening may be positioned at a second end of the serpentine chamber. The serpentine path may have a length extending from the first end to the second end. The length may be greater than the width and / or height of the serpentine chamber.

[0017] In some embodiments, the cryogenic storage container may include a hanger opening arranged to receive an IV bag hanger. The hanger opening may be positioned at an opposite end of the cryogenic storage container relative to the discharge opening such that when the cryogenic storage container is freely suspended from the IV hanger, the discharge opening is located at the lowest portion of the serpentine chamber. The hanger opening may be off-center along the end of the cryogenic storage container.

[0018] The cryogenic storage container of the present disclosure may include: a chamber having a volume; the chamber is defined by a wall portion of the cryogenic storage container, and wherein the wall portion of the cryogenic storage container each has an inner surface facing the chamber and an outer surface opposite to the inner surface; and wherein each inner surface has a surface area, and wherein the ratio of the sum of the surface areas to the volume of the chamber is between 1 to 5 (inclusive) and 2 to 5 (inclusive). The wall portion defining the chamber may follow a serpentine path from the first end of the chamber to the 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 between 5 mm (inclusive) and 7 mm (inclusive). The ratio may be between 3 to 10 (inclusive) and 2 to 5 (inclusive). In any of the disclosed arrangements, the ratio may be about 1 to 3.

[0019] The wall portion of the container may have a thermal conductivity between 0.16×10-3 Watts / Kelvin (inclusive) and 0.32×10-3 Watts / Kelvin (inclusive) at room temperature. In any of the disclosed arrangements, the wall portion may include a material having a thermal conductivity between 0.10 (inclusive) and 0.20 W / m·K (inclusive) 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 (e.g., metal) to increase the thermal conductivity of the material.

[0020] Advantageously, the exemplary cryogenic storage container shown in the provided figure can be oriented in multiple directions. For example, the cryogenic storage container can be positioned as a foot on a support surface, and the vent opening and the discharge opening of the cover are placed at the uppermost part of the cryogenic storage container. The cryogenic storage container can also be oriented with the discharge opening positioned at the lowest point of the chamber so that all liquids substantially contained in the cover chamber will be discharged through the discharge opening. In such an arrangement, the vent opening will also be positioned at the upper part of the chamber so as to allow air to enter the chamber without mixing with the discharged liquid and / or allow liquid to escape from the chamber through the vent opening. During the discharge process, the air entering the chamber through the vent opening is filtered by a filter associated with the vent opening to avoid contamination of the internal volume of the serpentine chamber.

[0021] Those skilled in the art will appreciate that cryogenic storage container can be formed by multiple manufacturing techniques, such as injection molding, blow molding, rotational molding, gas-assisted molding and rapid prototyping, to name a few non-limiting examples. Main body and lid can be manufactured separately, then such as by adhesive, ultrasonic welding, hot plate welding, infrared welding and / or laser welding attached together. In some manufacturing techniques (for example, rapid prototyping), lid and main body can be manufactured as a whole. The cryogenic storage container in the illustrated embodiment can be formed by at least injection molding and rapid prototyping. Preferably, no matter how manufacturing technology, container will be airtight sealable.

[0022] The cryogenic storage containers of the present disclosure preferably have rigid walls that maintain the same shape when filled or drained, regardless of the volume of the stored contents. Such arrangements can also provide a more consistent surface area to volume ratio for the fluid in the container compared to bags or other flexible storage containers. Advantageously, such arrangements can provide greater predictability and reliability during freezing and thawing cycles, which can help enhance cell viability.

[0023] Additionally, rigid containers are less prone to leaking than bags and, therefore, may be more preferred for situations where the stored contents are toxic or hazardous. One or more inner surfaces of the walls of the container (e.g., the inner surfaces of the first wall and the second wall) may be hydrophobic. Advantageously, such an arrangement may reduce cell adhesion.

[0024] Nucleation sites for ice crystal formation can be included on the inner surface of the wall defining the chamber. For example, the inner surface can include a coating with an ice nucleating agent (INA). Such a coating can be protected by a water-permeable top layer (e.g., titanium carbide, molybdenum disulfide with tungsten chalcogenide, and / or boron nitride with graphyne). By "seeding" ice formation, random formation of ice in the liquid during freezing can be reduced and / or eliminated, thereby improving sample survival. Random ice formation can occur at unpredictable temperatures; therefore, sample survival between repeated experiments with the same freezing protocol can be highly variable. In addition, extremely rapid crystallization results when ice forms in highly supercooled solutions, which can cause damage to cells and tissues. Nucleation sites provide control and increase the consistency of freezing distribution across samples and batches.

[0025] Advantageously, the arrangements disclosed herein can provide a cryogenic storage container having a chamber having a constant surface area / volume ratio along the length, width, and / or height of the chamber. For example, the chamber can have a volume defined by a wall portion of the cryogenic storage container, and wherein the wall portions of the cryogenic storage container each have an interior surface facing the chamber and an exterior surface opposite the interior surface, and wherein each interior surface has a surface area, and wherein the ratio of the sum of the surface areas to the volume of the chamber is between 1 to 5 (inclusive) and 2 to 5 (inclusive).

[0026] The containers of the present disclosure may also exhibit greater compressive strength than linear containers having the same wall thickness. Advantageously, the serpentine path of the sidewall of the container increases the compressive strength of the structure relative to a linear wall container in at least one direction.

[0027] The wall of the container may include one or more thermocouples. The thermocouples may be embedded in the wall. In some embodiments, one or more walls of the container may include one or more mounting features to facilitate consistent and reliable coupling of the thermocouple to the container. The thermocouple may be separated from the sample so as not to interfere with the nucleation behavior.

[0028] The container may include electrodes arranged to apply a high electric field in the kilovolt range for the purpose of electrofreezing and / or electroporation. Such electrodes may be coupled to one or more walls of the container.

[0029] The container may include a displacement body that protrudes into a chamber (e.g., a serpentine chamber) of the container. For example, one or more fins or rods may extend into the chamber and displace the volume of the sample therein. Advantageously, such an arrangement may further reduce the distance between a portion of the sample and the surface of the container. The one or more displacement bodies may be arranged to remove heat from the sample. For example, the one or more displacement bodies may have a thermal conductivity greater than the thermal conductivity of the wall of the container. As a specific example, the one or more displacement bodies may include metal and the container wall may include plastic. The thermocouple may also extend into the sample volume defined by the container wall.

[0030] The container may include an identification tag. Such a tag may include an electronic device (e.g., RFID, Bluetooth, etc.) for remote detection of the container and / or transmission of information from the electronic device. The container may include graduations indicating the volume of the sample in the chamber when the container is in a particular orientation.

[0031] The container may be optically transparent or otherwise include structures, features, and / or materials to facilitate visual inspection, optical quantitative measurements, and other types of non-destructive testing of the stored contents (e.g., spectroscopy, counting, fluorescence, etc.). In some embodiments, the container includes optical features similar to a cuvette.

[0032] The method for using the container disclosed herein includes removing oxygen from the chamber and / or purging the chamber with nitrogen through one or more openings. Electrical, mechanical, optical and / or acoustic measurements are performed on the liquid level through one or more openings. INA, sensors (e.g., optical and / or electrical sensors, such as thermocouples), cryoprotectants and / or cells are introduced into the chamber through one or more openings. The container is placed in a cryogenic freezer and / or the container is placed in a cryopreserved cell thaw, such as the VIA Thaw CB1000 by GE Lifesciences.

[0033] The containers disclosed herein can be hermetically sealed with cells and / or cryoprotectants positioned in the chambers.The containers disclosed herein can be provided in sealed sterile packaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is a perspective view of a cryogenic storage container according to an embodiment of the present invention.

[0035] Figure 2 yes Figure 1 A perspective view of the body of a cryogenic storage container.

[0036] Figure 3 yes Figure 1 Side view of a cryogenic storage vessel in a suspended / discharged configuration.

[0037] Figure 4 yes Figure 1 Side view of a cryogenic storage container in a filled configuration.

[0038] Figure 5 yes Figure 1 Side view of a cryogenic storage vessel in a discharge configuration.

[0039] Figure 6 , Figure 7 and Figure 8 yes Figure 1 A perspective view of the body of a cryogenic storage container.

[0040] Fig. 9 and Fig.10 yes Figure 1 A cross-sectional view of the body of a cryogenic storage container.

[0041] Fig.11 and Fig.12 yes Figure 1 A perspective view of a cover of a cryogenic storage container.

[0042] Fig.13 is a perspective view of a chamber for a cryogenic storage container according to another embodiment of the present invention.

[0043] Fig.14 is a perspective view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0044] Fig.15 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0045] Fig.16 is a perspective view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0046] Fig.17 yes Fig.16 A partial cross-sectional view of a chamber.

[0047] Fig.18 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0048] Fig.19 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0049] Fig. 20 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0050] Fig.21 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0051] Fig. 22 yes Fig.21 Detailed cross-sectional view of the chamber.

[0052] Fig.23 is a cross-sectional view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0053] Fig.24 is a perspective view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0054] Fig.25 is a perspective view of a chamber for a cryogenic storage container according to yet another embodiment of the present invention.

[0055] Fig.26 yes Fig.25 A partial cross-sectional view of a chamber.

[0056] Fig. 27 is a partial perspective view of a cryogenic storage container according to yet another embodiment of the present invention.

[0057] Fig.28 yes Fig. 27 A schematic partial cross-sectional view of a cryogenic storage container and a nucleation trigger.

[0058] Fig.29 yes Fig.28 Another schematic partial cross-sectional view of a cryogenic storage container and a nucleation trigger.

[0059] Fig.30 is a partial perspective view of a cryogenic storage container according to yet another embodiment of the present invention.

[0060] Fig.31 yes Fig.30 A partial perspective view of a cryogenic storage container and a thermocouple.

[0061] Fig.32 is a perspective cross-sectional view of a cryogenic storage container according to yet another embodiment of the present invention.

[0062] Fig.33 A perspective view of the interior side of a lower wall of a cryogenic storage container according to an embodiment of the present invention is shown.

[0063] Fig.34 Shows Fig.33 A perspective view of the exterior side of the upper wall of a cryogenic storage container.

[0064] Fig.35 A front view of the interior side of a lower wall of a cryogenic storage container according to an embodiment of the present invention is shown.

[0065] Fig.36 Shows Fig.35 A plan cross section of a cryogenic storage vessel. DETAILED DESCRIPTION

[0066] For the purpose of enhancing the understanding of the principles of the present invention, reference will now be made to the embodiments shown in the accompanying drawings, and specific language will be used to describe the embodiments. However, it should be understood that it is not intended to limit the scope of the present invention thereby. Any changes and further modifications in the described embodiments and any further application of the principles of the present invention as described herein are contemplated as would normally be expected by a person skilled in the art to which the present invention relates. An embodiment of the present invention is shown in considerable detail, and although it is apparent to a person skilled in the art to which the present invention relates, some features not relevant to the present invention may not be shown for the sake of clarity.

[0067] Except for terms expressly defined below, the language used in the claims and written description shall have only its plain and ordinary meaning. Such plain and ordinary meanings are defined herein to include all consistent dictionary definitions from the most recently published (as of the filing date of this document) general Merriam-Webster dictionary.

[0068] The terms "fill", "filling" and "filled" used herein include both partial filling and complete filling. The terms "drain", "draining" and "drained" used herein include both partial discharge and complete discharge. The term "vent" used herein refers to an opening that allows the balance of air pressure between the inside and outside of a cryogenic storage container. The vent may allow gas to pass through the vent (e.g., through a filter) and / or include a flexible membrane (e.g., a bag and / or a balloon) that allows expansion and / or contraction of gas in the chamber of a cryogenic storage container. The term "chamber width" used herein refers to the distance between the inner side surfaces on the opposite sides of the chamber, which is measured along a vector (i.e., vertical) perpendicular to at least one of the inner side surfaces. "Tubes", "tubings" and "flexible tubes" disclosed herein may include materials suitable for cryogenic storage at a temperature of -196°C or lower. Further forms, objects, features, aspects, benefits, advantages and embodiments of the present invention will become apparent from the detailed description and drawings provided herein.

[0069] With respect to the specification and claims, it should be noted that the singular forms "a," "an," "the," etc. include plural referents unless explicitly discussed otherwise. By way of illustration, reference to "a device" or "the device" includes one or more of such devices and their equivalents. It should also be noted that directional terms such as "up," "down," "top," "bottom," etc., are used herein merely for the convenience of the reader to aid the reader in understanding the illustrated embodiments, and the use of these directional terms is not intended to limit the described, illustrated, and / or claimed features to a particular direction and / or orientation in any way.

[0070] The terms "approximately" and "about" when qualifying a quantity, size, or geometry shall mean a tolerance of plus or minus 10% of a given value unless otherwise specified.

[0071] Unless otherwise indicated, all ranges disclosed herein encompass any and all subranges contained therein. For example, a range of "less than 10" may include any and all subranges between a minimum value of 0 and a maximum value of 10 (and including these two values), i.e., having a minimum value equal to or greater than 0 and a maximum value equal to or less than 10, e.g., 1 to 5. In certain cases, the numerical values ​​as described for the parameters may take negative values. In this case, the example values ​​of the range described as "less than 10" may take negative values, e.g., -1, -2, -3, -10, -20, -30, etc.

[0072] Figure 1 A cryogenic storage container 100 is shown including a body 102 . Figure 2 is a perspective view of the main body 102 . Figure 6 , Figure 7 and Figure 8 is a perspective view of the body 102 of the cryogenic storage container 100 . Fig. 9 and Fig.10 1 is a cross-sectional view of the main body 102 of the cryogenic storage container 100. Figure 1 , Figure 2 and Figures 6 to 10 , the body 102 defines a chamber 110 for containing 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 can also be defined by a first end wall 130 positioned at a first end 132 of the chamber 110 and a second end wall 134 positioned at a second end 136 of the chamber 110. A bottom wall 142 extends along and closes the bottom 140 of the chamber 110. The walls can be rigid to facilitate retention of shape.

[0073] 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 that are positioned at different angles relative to each other, or move from the first end 132 to the second end 136 at alternating angles. Along the serpentine path, there is a uniform characteristic dimension between the first wall 120 and the second wall 124, which is a uniform distance 156 between the first wall and the second wall measured along a direction perpendicular to the inside surface 150 of the first wall 120. The uniform distance 156 is uniform across 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 inside surface 150 of the first wall 120, can be between 5 mm (inclusive) and 7 mm (inclusive). The first wall 120 can be parallel to the second wall 124 along the serpentine path.

[0074] The first wall 120 and the second wall 124 may also have a uniform distance 156 between them along the height of the body. Thus, the first wall 120 and the second wall 124 may be separated by the uniform distance 156 across the length L and / or the height H. Advantageously, a uniform characteristic dimension is provided between the first wall 120 and the second wall 124 along the length L and / or the height H of the body 102, wherein the value of the uniform characteristic dimension is relatively small, which helps maintain a substantially uniform freezing distribution along the length L and / or the height H of the body 102.

[0075] In practice, the exact uniformity of uniform feature size is difficult to manufacture, so some tolerances are acceptable. In addition, in some embodiments, notches, 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 size on a portion of the wall of chamber 110. Therefore, in some embodiments, the range of uniform feature size 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 range of uniform feature size may, for example, deviate from the target value or intermediate value of the uniform feature size by 0.1%, 0.5%, 1%, 2%, 5%, 10%, 20% or 25%. In general, it is desirable that the higher uniformity of uniform feature size is to increase the uniform cooling and / or thawing rate of the contained biological sample / material.

[0076] In some embodiments, the uniform feature size may be uniform within any of the above tolerance ranges for 90% or more, 95% or more, or 99% or more of the perpendicular distance between the first wall 120 and the second wall 124 .

[0077] In some embodiments, the walls may be slightly separated from each other in a direction extending from the bottom 140 of the body 102 to the top 160 of the body 102. This separation may facilitate releasing the body 102 from the mold during manufacturing while not having a significant effect on changing the freezing profile along the height H of the body 102.

[0078] Advantageously, a serpentine chamber having a width, height, and overall length between the inner walls can provide a larger volume for liquid storage than a linear chamber having the same width, height, and overall length. Thus, a serpentine chamber arranged for a cryogenic storage box can store more liquid than a linear container of similar size (i.e., having an equal overall length).

[0079] The thermal resistance of the first wall 120 and the second wall 124 is preferably uniform along the length L and the height H. For example, the first wall 120 and / or the second wall 124 may have a uniform material thickness along the length L and the height H. The first wall 120 and the second wall 124 may have the same thermal resistance to promote uniform heat transfer during the freeze / thaw cycle.

[0080] Additional references Figure 3 , the cryogenic storage container includes one or more openings arranged to allow access to portions of chamber 110. For example, a first opening 202 of the cryogenic storage container may be in communication with first end 132 of chamber 110, a second opening 204 may be in communication with second end 136 of chamber 110, and a third opening 206 may be in communication with either first end 132 or second end 136 of chamber 110.

[0081] refer to Figures 1 to 3 , one or more of the openings 202, 204, 206 can be associated with a connector 207 (e.g., a Luer fitting or a barb fitting) that is arranged to couple to a liquid-filled 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 fit the resistance and / or interference within the flexible tubing. The liquid-filled device 212 is Figure 3 A flexible tube is shown connected to a barb fitting of opening 202. The flexible tube may be weldable (eg, 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 of the cryogenic storage container 100.

[0082] One or more of the openings 202, 204, 206 may be arranged for filling with liquid 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 arranged to drain air into and / or out of the chamber 110. For example, a vent opening arranged to drain air allows air to enter the chamber 110 when liquid is drained from the chamber 110, and a vent opening allows air to leave the chamber 110 when liquid is filled into the chamber 110. In the illustrated embodiment, a first opening 202 at a first end of the chamber 110 is arranged for venting, a second opening 204 at a second end of the chamber is arranged for draining, and a third opening 206 is arranged for filling the chamber with liquid.

[0083] In some embodiments, cryogenic storage container 100 may include a lid. Fig.11 and Fig.12 is a perspective view of the cover 300 of the cryogenic storage container 100. Figure 1 , Figure 2 , Fig.11 and Fig.12 , the cover 300 may close an end or side of the chamber 110. For example, in the illustrated embodiment, an upper portion of the chamber 110 defined by the top 160 of the body 102 is closed by the cover 300. The cover 300 may include one or more of the openings 202, 204, 206.

[0084] The bottom side 302 of the cover 300 may include a wall 310 extending between the vent opening (e.g., the first opening 202) and the fill opening (e.g., the third opening 206). The wall 310 prevents fluid entering through the fill opening from being immediately drawn out of the vent opening, particularly when negative pressure is applied to the vent opening to draw gas out of the chamber 110.

[0085] refer to Figure 3, filter element 220 can be associated with vent opening, and can be arranged to filter the air entering chamber 110 when chamber 110 is discharged.Filter element 220 can be a microfilter, more specifically a filter providing a microbial barrier.Such microfilter can be, for example, 3 μm, 0.2 μm or 0.1 μm sterile porous microfilter.In other embodiments, microfilter can have different structures.Filter element 220 can be arranged to be breathable but generally hydrophobic (that is, prevent the passage of liquid sample or cell suspension stored in container).Therefore, filter element 220 can retain suspension in cryogenic storage container 100, and when cryogenic storage container 100 is filled (partially or completely), gas is discharged from cryogenic storage container 100, and when suspension is extracted from cryogenic storage container 100, gas is discharged into cryogenic storage container 100. The filter element 220 is shown as being positioned / retained within the flexible tube 212 associated with the first opening 202, however the filter element 220 may otherwise be disposed in the air path through the vent opening.

[0086] The cryogenic storage container may include a suspension element 230, which may be a ring, such as Figures 1 to 3 20. The cryogenic storage container 100 may be suspended from the suspension element 230, as shown, or may be another arrangement now known or developed in the future for suspending the cryogenic storage container above a surface. The suspension element 230 may be arranged such that when the cryogenic storage container 100 is suspended from the suspension element 230, as shown in FIG. Figure 3 As shown, the discharge opening (e.g., opening 204) is located at the lowest point of chamber 110. Therefore, the fluid will flow toward the discharge opening under gravity. Such an arrangement can facilitate the discharge of fluid from chamber 110 to the greatest extent or completely, except for some residual fluids (such as moisture or fluid film) that may adhere to the inner surface of chamber 110. For example, the suspension element can be placed on the end opposite to the discharge opening and / or closer to the side opposite to the side with the discharge opening. The suspension element 230 can be offset from the center along the end of the cryogenic storage container 100.

[0087] The cryogenic storage container 100 can be arranged to support itself on a horizontal surface in a variety of positions. For example, the cryogenic storage container 100 can be arranged to support itself in a filled configuration. Figure 4 1 is a side view of a cryogenic storage container 100 in a filled configuration. In the filled configuration, the vent opening is elevated relative to at least 80% of the chamber. Thus, during filling, liquid rises toward the vent opening and gas in the chamber is exhausted through the vent opening. The cryogenic storage container 100 may also be arranged to be in a vented configuration (see Figure 3 and Figure 5 ) supports itself in a discharge configuration which may be the same as or different from the filling configuration. In the discharge configuration, the discharge opening is positioned at least 80% below the chamber.

[0088] In order to support itself in one or more configurations (e.g., filled and / or drained configurations), the cryogenic storage container 100 may include one or more brackets. Figure 2 , Figure 4 and Figure 5 The illustrated embodiment of the cryogenic storage container 110 has a first bracket 240 located at the first end 132 and a second bracket 242 located at the second end 136. In the filled configuration, Figure 4 As shown, the first bracket 240 and the second bracket 242 are in contact with the support surface 244, and the vent opening and the discharge / fill opening are raised relative to the chamber 110. In the discharge configuration, Figure 5 As shown, the cryogenic storage container 100 is supported on a support surface 244 by a second support 242, and the discharge opening is positioned below the chamber. The inner side surface of the lowest wall defining the chamber 110 (e.g., Figure 5 and Figure 8 The second end wall 134 shown is inclined downwardly toward the discharge opening in the discharge configuration and / or when the cryogenic storage container 100 is supported by the suspension element 230 so that substantially all of the fluid, except for possible residual fluid (e.g., <1% by volume), can be removed from the chamber 110.

[0089] In some embodiments, the cryogenic storage container 100 may 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 container 100 has a volume in the range of 100 ml to 250 ml.

[0090] As briefly described above, a relatively large capacity cryogenic storage container according to the present invention is provided with uniform feature dimensions and a high aspect ratio to promote consistency and uniformity of freezing / thawing rates. Such a container may have a serpentine chamber, such as the cryogenic storage container 100 described above. Alternatively, such a container may have other shapes. Several exemplary shapes of chambers of such a container are shown in Figures 13 to 26 and Figure 32 to Figure 34 was displayed in.

[0091] Reference now Fig.13 , a low temperature storage chamber 400 having a three-dimensional rectangular shape is shown. Chamber 400 has a uniform characteristic dimension R, more specifically a thickness dimension. Chamber 400 also has a height dimension H and a width dimension W. The aspect ratio for chamber 400 is defined as follows:

[0092]

[0093] The relationship between the surface area and aspect ratio of chamber 400 is as follows:

[0094]

[0095] For relatively high aspect ratios or relatively high ratios of SA1 to SA2 or SA1 to SA3, the characteristic dimension R significantly affects the freezing distribution of chamber 400 and provides a substantial uniformity of the freezing distribution. Such relatively high aspect ratios may be, for example, between 1:1 and 2:1, preferably between 2:1 and 5:1, and more preferably between 5:1 and 20:1.

[0096] refer to Fig.14 , a cryogenic storage chamber 500 having a cylindrical shape is shown. Chamber 500 has a uniform characteristic dimension R, more specifically a diameter. Chamber 500 also has a height dimension H. The aspect ratio for chamber 500 is defined as follows:

[0097]

[0098] The relationship between the surface area and aspect ratio of chamber 500 is as follows:

[0099]

[0100] For relatively high aspect ratios or relatively high ratios of SA1 to SA2, the characteristic dimension R significantly affects and provides substantial uniformity of the freezing distribution of chamber 500. Such relatively high aspect ratios may be, for example, between 1:1 and 2:1, preferably between 2:1 and 5:1, and more preferably between 5:1 and 20:1.

[0101] refer to Fig.15 , a cryogenic storage chamber 600 having a three-dimensional hollow cylindrical shape or a torus shape is shown. The chamber 600 has a uniform characteristic dimension R, more specifically a difference between an outer radius TA and an inner radius ra.

[0102] refer to Fig.16 and Fig.17 , a cryogenic storage chamber 700 having a plurality of connected three-dimensional square X shapes 702 is shown. Fig.17 , chamber 700 has a uniform characteristic dimension R, more specifically the vertical width of each lower limb of the X shape 702. Chamber 700 also includes a total thickness T, a total width W, and the number n of X shapes 702. Because the X shape 702 is a square, W / n=T. The lower limb of the X shape 702 has a pointed end 704. In some embodiments, the optimal value of R / T provides a relatively large perimeter for the chamber 700 without excessively reducing the area of ​​the chamber 700. In some embodiments, the value of R / T is in the range of 0.4 to 0.5, more specifically about 0.45. In some embodiments, chamber 700 has a height of about 80 mm, a total width W of about 120 mm, a total thickness T of about 20 mm, and a volume of about 103 ml.

[0103] refer to Fig.18 , a cryogenic storage chamber 800 is shown having a plurality of connected three-dimensional square X shapes 802. Chamber 800 is identical to chamber 700 described above, except that the lower limbs of X shapes 802 have hemispherical ends 804.

[0104] refer to Fig.19 , a cryogenic storage chamber 900 is shown 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.

[0105] refer to Fig. 20 , a cryogenic storage chamber 1000 having a three-dimensional rectangular shape with a plurality of cylindrical voids 1002 is shown. The chamber 1000 includes a total thickness T, a total width W, and a number of segments n. The chamber 1000 has uniform feature dimensions (not labeled), more specifically a vertical width between the cylindrical voids 1002.

[0106] refer to Fig.21 and Fig. 22 , a cryogenic storage chamber 1100 having a plurality of connected three-dimensional hollow hexagonal shapes 1102 is shown. Specifically referring to Fig. 22 , the chamber 1100 has a uniform characteristic dimension R, more specifically a vertical width of each of the hollow hexagonal shapes 1102. The chamber 1100 also includes a total thickness T, a total width W, and a number n of the hollow hexagonal shapes 1102.

[0107] refer to Fig.23 , a cryogenic storage chamber 1200 is shown having a plurality of three-dimensional hollow cylinders 1202. The chamber 1200 includes a total thickness T, a total width W, and a number n of hollow cylinders 1202.

[0108] refer to Fig.24 , a cryogenic storage chamber 1300 having nested hollow hexagonal shapes is shown. More specifically, chamber 1300 includes an outer hollow hexagonal shape 1302, an inner hollow hexagonal shape 1304, and a connector 1306 connecting the hollow hexagonal shapes 1302, 1304. Chamber 1200 may 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.

[0109] refer to Fig.25 and Fig.26 , a cryogenic storage chamber 1400 having a plurality of cylinders 1402 is shown. Specifically referring 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 .

[0110] refer to Figure 27 to Figure 29 , a cryogenic storage container 1500 is shown, which can generally have the same structure and dimensions as any of the cryogenic storage containers described herein. In addition, the cryogenic storage container 1500 includes one or more nucleation sites 1502 (illustratively, two nucleation sites) that promote the formation of ice crystals. Illustratively, each nucleation site 1502 is formed as a notch or recess 1504 in a wall 1506 of the cryogenic storage container 1500. In some embodiments, a method of freezing a sample facilitated by the nucleation sites 1502 is as follows. First, a sample 1508 is delivered to the cryogenic storage container 1500. The sample 1508 is then cooled (e.g., via placement in a controlled rate freezer) to a freezing point or below the freezing point (i.e., a supercooled state in which the water remains liquid but is very close to the freezing point via random ice nucleation). And then specifically as follows Fig.28 As shown, a nucleation trigger 1510 (e.g., a metal object cooled to below freezing) moves toward one of the nucleation sites 1502. Fig.29 As shown, nucleation trigger 1510 then contacts nucleation site 1502. Nucleation trigger 1510 thereby causes ice located at nucleation location 1502 to nucleate, and the ice nucleus then spreads outward from nucleation site 1502.

[0111] refer to Fig.30 and Fig.31 , a cryogenic storage container 1600 is shown, which can generally have the same structure and dimensions as any of the cryogenic storage containers described herein. In addition, one or more walls 1602 of the container 1600 may include one or more mounting features 1604, each of which receives a thermocouple 1606 ( Fig.31 ). The mounting feature 1604 prevents the thermocouple 1606 from being disengaged from the container 1600, which can cause temperature deviations in the manufacturing records. In addition, the mounting feature 1604 promotes a consistent location for temperature measurement across samples and batches. Illustratively, the mounting feature 1604 is formed as an external recess or channel 1608 in the wall 1602 and a wire retaining element 1610.

[0112] refer to Fig.32, cryogenic storage container 1700 is shown, and it can have the same structure and size as any one of the cryogenic storage containers described herein in general.Container 1700 includes relative upper wall 1702 and lower wall 1704 that limit chamber 1705.In this example, as described above, uniform characteristic dimension D1 can be changed to include one or more recesses for specific purposes other than promoting consistent and uniform freezing / thawing rate.In this example, cryogenic storage container 1700 promotes visual inspection, optical quantitative measurement and other types of non-destructive testing of the contents of container 1700.Relative upper wall 1702 and lower wall 1704 can be transparent or include transparent parts that are convenient for microscopy and spectroscopy.Such process in turn promotes measurement, diagnosis and early identification in the process of sample problems.In certain embodiments, upper wall 1702 and / or lower wall 1704 include one or more recesses to prevent the destruction of corresponding wall 1702,1704 during manufacture, distribution and / or use.

[0113] Illustratively, the upper wall 1702 includes one recess 1706 and the lower wall 1704 includes two recesses 1708. In some embodiments, the upper wall 1702 and the lower wall 1704 define an optical path length 1710 therebetween, and the optical path length 1710 can be uniform across a set of containers of different volumes. The one recess 1706 and the two recesses 1708 can be sufficiently deep to meet their intended purpose of reducing or preventing damage to the walls 1702, 1704, while also being sufficiently shallow to maintain sufficient uniformity of uniform feature dimension D1 (e.g., within the tolerances described above) and sufficient consistency / uniformity of freezing / thawing rates in the chamber.

[0114] Fig.33 The exterior side of the upper wall 1802 of the cryogenic storage container 1800 is shown, and Fig.34 The interior side of lower wall 1804 of cryogenic storage container 1800 is shown. Fig.33 and Fig.34 Also shown are vent line 1820, fill line 1822, and tip port 1824. Similar to Fig.32 In this example, the uniform feature size (not shown, but similar to Fig.33D1 of the embodiment of the present invention is comparable) due to the inclusion of one or more recesses for a specific purpose other than promoting a consistent and uniform freezing / thawing rate, the specific location or area between the upper wall 1802 and the lower wall 1804 can be changed. The upper wall 1802 includes a first recess 1806 and the lower 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 can be deep enough to meet their intended purpose, while also being shallow enough to maintain sufficient uniformity of uniform feature size (e.g., within the tolerances discussed above) and sufficient consistency / uniformity of the freezing / thawing rate in the chamber. For example, the uniform feature size or vertical distance between the upper wall 1802 and the lower wall 1804 can be a maximum of 10 mm without a recess and a minimum of 8 mm with a recess in each wall 1802, 1804. Thus, the uniform feature size can be considered to be 10 mm, with a deviation of minus 20% in the vertical distance between the upper wall 1802 and the lower wall 1804 extending between the two recesses. In this example, the maximum height H of the cryogenic storage container 1800 is approximately 100 mm, making the aspect ratio approximately 10 to 1.

[0115] Fig.35 The interior side of the lower wall 1904 of the cryogenic storage container 1900 is shown, and Fig.36 A cross section of a plane parallel to uniform characteristic dimension D2 is shown for cryogenic storage container 1900. Cryogenic storage container 1900 is similar to cryogenic storage chamber 1800, but has an upper wall 1902 with two recesses, and has the recesses in a different shape.

[0116] Although the present invention has been shown and described in detail in the drawings and the foregoing description, these embodiments should be considered illustrative rather than restrictive in nature, and it should be understood that only preferred embodiments have been shown and described, and it is intended to protect all changes, equivalents and modifications within the spirit of the present invention as defined by the following claims. All publications, patents and patent applications cited in this specification are incorporated herein by reference, as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference and is fully listed herein.

Claims

1. A cryogenic storage container, comprising: A first body defining a chamber, the first body comprising a plurality of walls, the plurality of walls comprising: a first wall extending along a first side of the chamber; and a second wall extending along a second side of the chamber, the second side being opposite the first side relative to the chamber, The chamber has a volume in the range of 25 mL to 250 mL, the chamber having a uniform characteristic dimension extending perpendicularly to the first wall and the second wall from the first wall to the second wall, the uniform characteristic dimension being uniform across a planar cross-section of the chamber parallel to the uniform characteristic dimension. 2 . The cryogenic storage container of claim 1 , wherein the uniform characteristic dimension is uniform for 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. The cryogenic storage container according to claim 1, further comprising: a vent opening in fluid communication with the chamber; as well as A discharge opening is in fluid communication with the chamber.

4. The cryogenic storage container of claim 3, further comprising a filter associated with the vent opening to filter air entering and / or leaving the chamber.

5. The cryogenic storage container of claim 3, further comprising a heat-sealing tube attached to at least one of the group consisting of the discharge opening and the vent opening.

6. The cryogenic storage container of claim 3, wherein the vent opening is positioned at a first end of the chamber and the vent opening is positioned at a second end of the chamber. The cryogenic storage container of claim 1 , wherein the chamber follows a serpentine path.

8. The cryogenic storage container of claim 7, wherein the body further comprises: a first end wall at a first end of the chamber, the first end wall connecting the first wall and the second wall; as well as a second end wall at a second end of the chamber, the second end wall being opposite the first end wall relative to the chamber, Wherein the serpentine path extends along a length between the first end wall and the second end wall.

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

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

11. The cryogenic storage container of claim 7, wherein the serpentine path is defined by a series of straight wall sections arranged in a herringbone pattern.

12. The cryogenic storage container of claim 1, wherein the uniform characteristic dimension is between 5 mm (inclusive) and 10 mm (inclusive).

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

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 when the container is freely suspended from the IV bag hanger, the drain opening is located at a lowermost portion of the serpentine chamber.

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

16. The cryogenic storage container of claim 1, further comprising nucleation sites that promote ice crystal formation.

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

18. The cryogenic storage container of claim 1, wherein the plurality of walls include mounting features, and wherein the cryogenic storage container further includes a thermocouple carried by the mounting features.

19. The cryogenic storage container of claim 18, wherein the mounting feature comprises a recess.

20. The cryogenic storage container of claim 19, wherein the mounting feature further comprises a wire retention element.

21. The cryogenic storage container of claim 1, wherein the plurality of walls include transparent wall portions.

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

23. The cryogenic storage container of claim 1, further comprising: a second body defining a second chamber, the second body comprising 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 fourth wall extending along a second side of the second chamber, the second side of the second chamber being opposite to the first side of the second chamber relative to the second chamber, the second chamber having a second volume in a range of 25 mL to 250 mL, the second chamber having a second uniform characteristic dimension extending perpendicular to the third wall and the fourth wall from the third wall to the fourth wall, the second uniform characteristic dimension being uniform across a plane cross-section of the second chamber parallel to the second uniform characteristic dimension, The first chamber is in fluid communication with the second chamber.

24. The cryogenic storage container of claim 23, further comprising: a third body defining a third chamber, the third body comprising a third plurality of walls, the third plurality of walls comprising: a fifth wall extending along the first side of the third chamber; and a sixth wall extending along a second side of the third chamber, the second side of the third chamber being opposite to the first side of the third chamber relative to the third chamber, The third chamber has a third volume, the third chamber has a third uniform characteristic dimension extending perpendicularly to the fifth wall and the sixth wall from the fifth wall to the sixth wall, the third uniform characteristic dimension being uniform across a plane cross-section of the third chamber parallel to the third uniform characteristic dimension, The second chamber is in fluid communication with the third chamber.

25. A cryogenic storage container, comprising: a chamber, which has a volume; as well as wall portions defining the chamber, the wall portions each having an inner surface facing the chamber and an outer surface opposite the inner surface, Each interior surface has a surface area, and the ratio of the sum of the surface areas of the interior surfaces to the volume of the chamber is between 1 to 5 (inclusive) and 2 to 5 (inclusive).

26. The cryogenic storage container of claim 25, wherein the wall portion defining the chamber follows a serpentine path from a first end of the chamber to a second end of the chamber.

27. The cryogenic storage container 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. The cryogenic storage container of claim 27, wherein the width is between 5 mm (inclusive) and 10 mm (inclusive).

29. The cryogenic storage container of claim 25, wherein the wall portion has a relative humidity of 0.16×10- 3 Watt / Kelvin (inclusive) and 0.32×10- 3 Thermal conductivity in W / Kelvin (inclusive).

30. The cryogenic storage container of claim 25, wherein the wall portion comprises a material having a thermal conductivity of between 0.10 (inclusive) and 0.20 W / m·K (inclusive) at room temperature.

Images