Composite function cryogenic storage container

Abstract

A system and device for the storage of cryogenically preserved materials with improved stability to mechanical shock. The device comprises a container with an increased surface area to volume ratio when compared to a cylindrical container of comparable capacity, further comprising a material that provides a continuous barrier of a single material surrounding the container contents while maintaining resistance to shock and impact even at cryogenic temperatures. The device further comprises an internalized sensor and an enhanced interface with external equipment, and a design that is easily modifiable to provide optimized thawing rates and post-thaw sample processing capabilities.

Description

[Technical field]

[0001] The present invention relates to a vessel device that reliably contains frozen cell suspensions at cryogenic temperatures while providing improved protection from impact, shock and acceleration damage. The present invention also relates to a cryogenic storage vessel having sensors and a communication interface. Furthermore, the present invention relates to a cryogenic storage vessel that facilitates interaction between the vessel and other devices during storage, transportation, and during and after the thawing process. Finally, the present invention relates to a cryogenic storage vessel configured for post-thawing fluidic and cellular processing in containers stored within the vessel prior to removal of the vessel contents after thawing. [Background technology]

[0002] The field of live cell-based pharmacology is well established and will continue to expand. Many cell-based therapeutics require the expansion of cell populations to larger numbers through various processing steps. Once the cell population is expanded, the need to store the cells for extended periods of time often arises. For example, cells may need to be administered to a recipient for an extended period of time. It may be a therapy intended for administration to a group of as yet unidentified individuals, or it may be necessary to store the cell population indefinitely as a future resource or reference sample. Another exemplary reason may simply be that the recipient is in a remote location and the interval between shipments is too long. However, cells are living and dynamic. Simple storage is not possible for these organisms, as cells in culture continue to grow and require a fresh source of nutrients and a means of waste removal. However, there are means to metabolically arrest the time of cell suspensions, including suspending the cells in a cryogenic storage solution and freezing the cells at a defined rate of temperature loss. When the suspension solidifies and the temperature is sufficiently reduced, the frozen cells may be stored indefinitely, provided that the temperature of the frozen mass is kept below a defined maximum. Cell suspensions cryopreserved in this manner are typically stored in vacuum insulated storage vessels that use liquid nitrogen as the refrigerant, thus typically maintaining the frozen cells at a temperature range of -196°C to -150°C.

[0003] Currently commercially available cryogenic storage containers are divided into two main groups. The first group includes rigid containers, typically constructed from molded resin materials. Due to selected material properties and wall thickness specifications, these rigid containers have sufficient toughness to withstand forces and impact shocks at cryogenic temperatures. The geometric configuration of rigid containers is predominantly cylindrical, narrowed at the access port. To provide a means for filling and dispensing the contents, the port opening is typically plugged with a septum plug of a material such as rubber that can be easily penetrated by a hypodermic needle or a specially designed filling needle. Due to the difficulty of joining the rubber material to the resin material with a joint that remains intact at cryogenic temperatures, as well as the difference in expansion rates between the vial resin material and the septum material, the rubber septum plug is often secured to the rim of the resin vial by a secondary retainer of aluminum or plastic. While the retainer is effective in preventing separation and can provide additional clamping force, the hardening and shrinkage of the rubber material at temperatures below -50°C can make the container seal unreliable at cryogenic temperatures, thereby raising concerns regarding contamination, release of live material into the environment, and explosive release of the rubber plug after liquid nitrogen aspiration.

[0004] Additional challenges posed by rigid containers come from the difficulty of interfacing with a solid heating block due to the curvature of the container. Also, in cylindrical containers, the volume to surface area ratio decreases with increasing radial dimensions as the volumetric capacity increases, which limits the heat inflow rate during the melting process and increases the melting interval. In a rigid container during thawing, when thermal energy is introduced into the container from the inner wall boundary, phase transformation begins at the inner wall and progresses to the axis of the cylindrical frozen mass. Thus, during the thawing process, a thermal gradient is introduced, which exposes cells in the gradient region to temperatures that may damage the cells for an extended period of time.

[0005] The second group of storage containers includes flexible containers, such as those in the form of flexible cryopreservation bags. While these types of storage containers offer potential advantages over rigid-walled containers, they introduce vulnerability to motion damage when compared to rigid-walled containers. Cryopreservation bags are typically not filled to their maximum capacity, but rather to a capacity that allows for a typical thickness limit of about 1 cm. The limited thickness of the frozen mass combined with the large surface area of ​​the bag allows for greater control of the cooling rate during the freezing process, as well as significantly reducing the intervals during the thawing process compared to rigid containers for comparable content volumes. Additionally, the flexibility of the bag material allows for convective distribution of thermal energy during the thawing process due to movement of the container walls and mixing of the contents.

[0006] The primary vulnerability of cryopreservation bags is their susceptibility to fracture at low temperatures. Cryopreservation bags are generally constructed of a laminate of two sheets of polymeric material, the boundary edges of which are joined by a fused heat seal. The stiffness of the polymeric sheet material increases such that it becomes brittle as the temperature of the material becomes cryogenic. As a result, impacts to the cryopreservation bag, especially along the boundary edges, can result in crack formation or fracture of the bag material. Contents of the cryopreservation bag are typically introduced and withdrawn through a port assembly fused between the primary laminated layers along the seam edge. Protrusion of the port assembly imposes the potential for impact or torsional forces that concentrate stresses leading to failure at the port-bag junction. Although ports provide a more reliable continuous material seal than those constructed from multiple joined materials, the seal is typically pierced using a tube needle or spike to remove the contents of the container, so the potential risk of accidentally puncturing the bag wall is a concern unless specific protective design features are incorporated.

[0007] A further drawback of cryopreservation bags comes from the aspect that when the liquid contents are withdrawn, the inner walls of the bag are placed close together such that, even with the aid of gravity, the surface tension of the aqueous solution acts to retain the liquid within the bag, resulting in a loss of dead volume that cannot be recovered unless special methods are applied, such as rolling the bag.

[0008] Thus, while various methods and devices currently exist for the storage of sensitive frozen materials, several challenges remain. The devices and methods of the present invention address and meet these needs. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention relates generally to devices that provide storage for liquid samples that are subsequently cryopreserved for cryopreservation purposes. [Means for solving the problem]

[0010] In a first aspect, the present invention is directed to a cryogenic storage container that provides protection from impacts, shocks, and acceleration or damage due to applied mechanical forces, where when the storage container is filled and sealed, the storage container comprises a complete and continuous interior surface of a single material joined at the seam closed by fusion bonding. In some embodiments, the container is constructed from a cryogenic compatible material that provides toughness to reduce embrittlement and maximizes flexibility and resistance to impact and stress fracture at cryogenic temperatures, while being sufficiently flexible and pliable at ambient and aqueous freezing temperatures to allow limited adjustment of volume capacity by compressive mechanical forces or by expansion pressure of the aqueous contents upon solidification. In some embodiments, the present invention provides a cryogenic storage container that is semi-rigid such that the dimensions of the container do not collapse in a gravitational field, thereby preventing internal surface tension forces that would result in retention of liquid contained therein when emptied.

[0011] In some embodiments of the invention, varied dimensions are provided to control the volume to surface area ratio of the container. For example, in some embodiments, one dimension of the container is minimized while the remaining two dimensions are adjusted to optimize the planar area on the two major parallel planes, thereby controlling the volume to surface area ratio such that contact of the larger planar surface with an external planar heat sink or heat source provides a rapid transition time between the cryogenic temperature and the melting temperature of the container contents. To allow expansion of the aqueous contents during freezing and provide an option to minimize air voids while filling the container, some embodiments of the invention provide one or more rim expansion curvatures that allow an increase in the separation distance of at least two major container surfaces while maintaining the parallel plane orientation of the major container surfaces. In some embodiments, a first dimension of the container is made smaller than the remaining two dimensions of the container, where the two major dimensions control the area of ​​the major surface of each container shell portion, and where the third dimension controls the distance between the major surfaces of each shell portion when the container is assembled. In some embodiments, the ratio between the first dimension and the sum of the second and third dimensions is 1:1 to 4:1. In some embodiments, the ratio between the first dimension and the third dimension is from 1:1 to 30:1. In some embodiments, the ratio between the first dimension and the third dimension is from 2:1 to 20:1. In some embodiments, the container of the present invention comprises an area determined by the first dimension and the second dimension, where the area is less than 3 cm. 2 From 300cm 2 Up to 300cm 2 From 1000cm 2 That is until now.

[0012] In some embodiments, the storage container comprises two halves that each include a nominally rectangular major surface with rounded corners and that are substantially mirror images of one another, with the periphery of each major surface joined with a swept profile rim and the opposite or unjoined end of the swept profile rim joined with an outward flange, where the two container portions are configured to be seam joined at the mirror surfaces of the two halves with a fused flange seal. In some embodiments, the flange of the present invention comprises a width of greater than 1 mm. In some embodiments, the container comprises a first half or part having a nominally rectangular major surface with rounded corners, the periphery of the major surface being joined with a swept profile rim and an opposite or unjoined end of the swept profile rim being joined with an outward flange that is joined with a planar surface of a second half or part, where joining the first and second parts by a heat welded joint at the flange surfaces closes the interior of the container.

[0013] In some embodiments, the invention provides a container including a cavity in which one or more halves or portions of the container have a non-linearly swept rim wall profile configured to expand and contract to allow adjustment of an interior volume of the cavity, where the non-linearly swept rim wall profile flares outwardly from a major surface of the container portion toward a rim flange of the cavity.

[0014] In some embodiments, the melt flange and rim profile of the container may engage a bumper frame configured to protect the container during the product's life cycle, including filling, freezing, transport, storage and thawing procedures.

[0015] In some embodiments, the container includes one or more access ports located away from one or more midline seams, offset seams, or minor surfaces of the container, thereby protecting the access ports from impacts, shocks, or unintended mechanical strain or stress. In some embodiments, the access ports include integrated ports, where the integrated ports are an uninterrupted continuum of the container wall material, thereby providing an access port seal that does not rely on bonded seams of similar or dissimilar materials. In some embodiments, no additional construction of the access port is required beyond molding a particular recess or curvature in the container surface, while in other embodiments, access to the contents of the container may be achieved by directly penetrating the container wall. In some embodiments, the recesses that serve as access ports in the container surface may be occluded by a removable membrane, which maintains the sterility of the access port after sterilization of the completed container until the membrane is removed.

[0016] In a second aspect, the container includes a first half or portion having a nominally rectangular major surface with rounded corners, the periphery of the major surface being joined with a swept profile rim, the opposite or unjoined end of the swept rim being joined with an outward flange, the first portion being joined at the surface plane of the flange with a second half or portion including a planar surface, and the first and second portions being joined by a heat welded joint, thereby closing off the interior of the container. In some embodiments, the container further includes a driver element including a flat, thin sheet-like formation of ferrite material laminated between two layers of material having the same or similar properties as the material of the first and second portions of the container, such that when the driver element is disposed within the container, the driver element deflects the solid contents of the container to a specific region of the container's interior when exposed to an external magnetic field. In some embodiments, the container's planar wall includes an electronic circuit having circuit leads terminating at one or more locations on the end of the planar wall. In some embodiments, the container includes an electronic circuit having circuit leads terminating at one or more locations on the exterior surface of the planar wall. In some embodiments, at least one of the electronic circuits includes a sensor for measuring and / or monitoring one or more parameters of the contents of the container, such as temperature, pH, alkalinity, or oxygen level.

[0017] In a third aspect of the invention, a container is provided that includes an outer wall constructed from two opposing portions joined at a mid-planar seam and an additional seam forming a sealed division of the container, where one dimension of the container is smaller than the other two dimensions such that the container has two large planar surfaces joined by a swept rim wall that terminates in a flange seam joined by heat welding. The additional seam also joins the swept rim through an interior portion of the container such that the container is divided into two or more compartments that are completely isolated from each other, except at specific locations where a through duct allows communication between two separate enclosed compartments or volumes. In some embodiments, one or more of the through ducts are normally closed by a valve mechanism. In some embodiments, the valve opens when the container is subjected to sufficient centrifugal force adjusted to a specific vector. In some embodiments, a method of using the container of the invention comprises filling a first cavity of the container with a cell suspension containing a cryopreservation liquid and filling a second cavity of the container with an exchange medium, whereby upon thawing the cryopreservation medium, the cells migrate from the first cavity to the second cavity, whereby the cryopreservation medium is substantially diluted or absent. In some embodiments, the cells in the first cavity of the container are urged through one of the ducts by centrifugal force, at the same time that the same centrifugal force urges the opening of a duct communicating with the first and second cavities, thereby allowing the passage of the cells to the second cavity. In some embodiments, the second communicating duct connecting the first and second cavities is blocked by a valve located near the axis of rotation, whose angular displacement produces a centrifugal force on the container. At a sufficient level of relative centrifugal force, the valve of the second communicating duct is opened and proximate the remainder of the container, and the liquid contents of the container are displaced to the distal portion of the container, so that the open valve of the second communicating duct allows the gas pressure to equilibrate between the two container chambers, thereby eliminating the pressure difference between the two chambers in the absence of centrifugal force.

[0018] In a fourth aspect, the invention includes a method for using a container according to the first to third aspects above. Some aspects of the method apply to a fill and seal sequence that may be used to introduce a liquid into the container in a sterile manner and then seal the container with a molten seal. Other aspects of the method apply to a sequence of steps that may be used to aseptically extract the liquid contents from the container.

[0019] These aspects, embodiments and advantages of the present invention are explained in greater detail in the accompanying drawings and the detailed description that follows. [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 is an exploded view showing major elements of a container assembly according to an exemplary embodiment of the present invention. [Figure 2A] FIG. 2 is a perspective view of an assembled representation of the container assembly shown in FIG. [Figure 2B] FIG. 2B is a cross-sectional view of the assembly embodiment shown in FIG. [Figure 2C] FIG. 2B is a detailed cross-sectional view of the assembly embodiment shown in FIG. [Diagram 3] Dimensional diagram of the embodiment of the present invention shown in FIGS. 1 and 2A-2C [Figure 4A] FIG. 1 is a detailed perspective cross-sectional view of a port recess according to an exemplary embodiment of the present invention. [Figure 4B] FIG. 1 is a detailed perspective cross-sectional view of a port recess according to an exemplary embodiment of the present invention. [Figure 4C] FIG. 1 is a detailed perspective cross-sectional view of a port recess according to an exemplary embodiment of the present invention. [Figure 4D] FIG. 1 is a detailed perspective cross-sectional view of a port recess according to an exemplary embodiment of the present invention. [Figure 5A] FIG. 1 is a detailed perspective view of a shell storage container demonstrating a method of filling and sealing the shell storage container of the present invention, according to an exemplary embodiment of the present invention. [Figure 5B] FIG. 1 is a detailed perspective view of a shell storage container demonstrating a method of filling and sealing the shell storage container of the present invention, according to an exemplary embodiment of the present invention. [Figure 5C]FIG. 1 is a detailed perspective view of a shell storage container demonstrating a method of filling and sealing the shell storage container of the present invention, according to an exemplary embodiment of the present invention. [Figure 5D] FIG. 1 is a detailed perspective view of a shell storage container demonstrating a method of filling and sealing the shell storage container of the present invention, according to an exemplary embodiment of the present invention. [Figure 5E] FIG. 1 is a detailed perspective view of a shell storage container demonstrating a method of filling and sealing the shell storage container of the present invention, according to an exemplary embodiment of the present invention. [Figure 6A] 1A and 1B are perspective and cross-sectional views of an exemplary through-vessel sterile filter vent tool that may be used with the shell storage vessel of the present invention; [Figure 6B] FIG. 1 is a partial cross-sectional side view of a shell storage vessel in which the shell storage vessel has been penetrated by a sterile filter venting tool, according to an exemplary embodiment of the present invention, where this view demonstrates how the shell storage vessel is vented. [Figure 7] FIG. 13 is a detailed perspective cross-sectional view of a port recess having a threaded interface for mounting an external accessory welded element therein, according to an exemplary embodiment of the present invention; [Figure 8A] FIG. 1 is a detailed cross-sectional view of a shell storage vessel having a swept rim profile and a planar wall surrounding the vessel according to an exemplary embodiment of the present invention. [Figure 8B] FIG. 1 is a detailed cross-sectional view of a shell storage vessel having a swept rim profile and a planar wall surrounding the vessel according to an exemplary embodiment of the present invention. [Figure 9A] FIG. 1 is a perspective view of a shell storage container having a rim extension that allows the container to be filled through a channel that extends through a swept rim, according to an exemplary embodiment of the present invention. [Figure 9B] An exploded view of the embodiment shown in FIG. [Figure 9C] FIG. 9B is a perspective view of the embodiment shown in FIG. 9A during a first stage of container filling according to an exemplary embodiment of the present invention. [Figure 9D] FIG. 9B is a perspective view of the embodiment shown in FIG. 9A after a filling and sealing stage, according to an exemplary embodiment of the present invention. [Figure 9E]FIG. 9B is a perspective view of the embodiment shown in FIG. 9A sealed and with the fill access port cut away, according to an exemplary embodiment of the present invention. [Figure 9F] Dimensional drawing of the embodiment shown in FIG. [Figure 10A] FIG. 1 is an exploded view showing the main elements of an embodiment of the present invention, including an internal driver card receptacle solid content deflection element, having a planar receptacle wall adjacent a flange joint, according to an exemplary embodiment of the present invention. [Figure 10B] FIG. 10B is a perspective view of an assembled representative example of the embodiment shown in FIG. [Figure 11A] FIG. 10C is a cross-sectional view of the embodiment shown in FIG. 10B in contact with a cross-sectional view of an exemplary freezing device to illustrate an early stage of an exemplary method of use according to an exemplary embodiment of the present invention. [Figure 11B] 11A in cross section of the embodiment shown in FIG. 10A in contact with a cross section of an exemplary freezing device to illustrate a later stage of the example method of use shown in FIG. [Figure 11C] 10A in contact with a cross-sectional view of an exemplary freezing device to illustrate a later stage of the example method of use shown in FIG. 11B. [Figure 11D] 11C is a cross-sectional view of the embodiment shown in FIG. 10A in contact with a cross-sectional view of an exemplary freezing device to illustrate a later stage of the example method of use shown in FIG. [Figure 12] FIG. 1 is an exploded view showing the main elements of an embodiment of the present invention, including a solidification vessel contents deflection driver card and a sensor circuit embedded in a second vessel shell portion, according to an exemplary embodiment of the present invention. [Figure 13A] FIG. 1 is a perspective view of an embodiment of a vessel divided into two specific chambers connected by a valve that is actuated by an acceleration force, according to an exemplary embodiment of the present invention. [Figure 13B] FIG. 13B is an exploded view of the vessel shown in FIG. 13A revealing the internal chamber and valve structure according to an exemplary embodiment of the present invention. [Figure 14A] FIG. 1 is a detailed view of a centrifugally actuated gas valve subcomponent according to an exemplary embodiment of the present invention; [Figure 14B] Cross-sectional view of the gas valve shown in FIG. [Figure 15A] Detailed view of a centrifugally actuated cell gate valve according to an exemplary embodiment of the present invention. [Figure 15B] Cross-sectional view of the cell gate valve shown in FIG. [Figure 16A] FIG. 1 is a perspective view of a partially assembled shell storage vessel coupled to an exemplary centrifuge machine for applying centrifugal force to the exemplary embodiment, in accordance with an exemplary embodiment and method of the present invention; [Figure 16B] FIG. 16B is a cross-sectional view of the shell storage vessel of FIG. 16A fully assembled and installed in accordance with an exemplary embodiment of the present invention, further illustrating vessel shape limiting features of a centrifuge machine in accordance with an exemplary embodiment of the present invention. [Figure 16C] FIG. 13C is a perspective top view of a second lower shell portion of the shell storage container of FIGS. 13A and 13B; DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] The present invention provides a cryopreserved cell suspension storage container and storage system that meets many properties and functionalities not offered by currently available containers. In some embodiments, the container of the present invention provides a means for containing liquids and / or aqueous solutions, such as solutions, compositions, formulations, particulate suspensions, virus suspensions, cell suspensions, and multicellular organism suspensions. In some embodiments, the liquid is contained within the container by a barrier of a single material, where the various parts comprising the container are joined by melt bonding of the single material, which is not itself vulnerable to separation of parts and fracture of bonds under stresses imposed by exposure to low and cryogenic temperatures. In some embodiments, the single material has properties that provide self-supporting geometric rigidity and selective penetrability at ambient temperatures, while retaining toughness, flexibility, and resistance to fracture at cryogenic temperatures. In some embodiments, the single material is a thermoplastic polymer or polymer blend that can be vacuum formed from sheet material, thereby providing an inexpensive means of construction that is acceptable by low-cost vacuum forming and heat sealing, in addition to allowing low-cost thermoforming of essential local features within a part prior to final assembly. In other embodiments, the container may be manufactured from the same material using an injection molding system. In some embodiments, attachment and removal of the container contents may be accomplished by direct penetration of the container wall. In some embodiments, punctures in the container wall during the filling process may be sealed with a plug constructed from the same material as the container wall and sealed in place by heat sealing of the plug to the container wall. In some embodiments, a special fill port is provided that forms a continuum with the container wall, where the special fill port can be used to introduce the container contents and simultaneously or subsequently collapse sealed by a heat seal bond at or near the port-container wall junction with removal of the portion of the port structure distal to the heat seal bond or seal. In some embodiments, special port structures are molded into the surface of the container, where the ports are configured to facilitate the introduction or removal of the container contents by penetration of the container wall.In some embodiments, the special port structure or port bay is recessed from the major surface of the container such that no portion of the port structure protrudes beyond the major surface of the container. In some embodiments, the recessed port structure or bay is occluded by a removable hermetic seal that maintains sterility of the recessed port bay volume until removed. In some embodiments, the storage container comprises an enclosure of the same material that forms a continuous wall barrier at all points of contact with the contents of the container. In some embodiments, the container is constructed from two parts, and in other embodiments, the container is constructed from more than two parts. In some embodiments, the wall of the container is constructed from two shell parts, each with a rim flange, and the two shell parts are joined at their respective rim flanges via a heat fusion bond. In some embodiments, the rim flange comprises one surface that is entirely planar. In some embodiments, at least one rim flange deviates from a planar configuration in one or more locations. In some embodiments, the two shell portions of the container are symmetrical with respect to a plane located along the fusion bond of the two shell portions. In some embodiments, the two shell portions that form the shell of the container are asymmetrical. In some embodiments, one of the container shell portions or halves is planar.

[0022] In some embodiments, the dimension of the container perpendicular to the heat-sealed flange joint is shorter than the other two dimensions such that the container is nominally in the shape of a flattened rectangle. In some embodiments, the flattened rectangular shape includes two major planes and parallel faces on opposite sides of the flange joint. In some embodiments, the container wall intervening between the planes and the respective flanges on the same portion includes a swept rim profile. In some embodiments, the swept rim profile includes a curvature that allows the dimension between the flange and the plane to decrease or increase at both ambient and lower temperatures, where a phase change of the aqueous contents of the container occurs at the lower temperature.

[0023] In some embodiments, the molded port bay is located on a surface of the container such that the port bay does not occupy any part of the heat seal bond, does not participate in the heat seal bond, or is not an intervening structure in the heat seal bond that joins the two shell parts of the container exterior after the container contents are attached. In some embodiments, the port bay is confined to one of the two shell parts that form the container. In some embodiments, the port bay may be located in one or more of the shell parts. In some embodiments, none of the shell parts of the container include the port bay. In some embodiments, the port bay may be molded into the rim wall of the container. In some embodiments, selected regions of the port bay wall may be reduced in thickness to a particular dimension by localized molding and / or thermoforming methods, where penetration of the container wall may be facilitated for installation and removal of the container contents. In some embodiments, selected regions of the port bay may be thermally altered to form a recess or well shape in the port bay wall. In some embodiments, the recess includes a single radially symmetric wall. In some embodiments, the recess includes at least two walls. In some embodiments, at least two walls include a partial region having a thickness less than the wall thickness of the remaining portions of the at least two walls. In some embodiments, the recess is located on an edge of at least one of the first and second major surfaces of the first or second shell of the container of the present invention. In some embodiments, the recess is located on a swept rim of the container of the present invention.

[0024] In some embodiments, the flange rim includes an extension at one or more locations on or along the flange, the extension forming a conduit that terminates at the vessel access port. In some embodiments, the access port includes a septum plug or plug assembly of a rubber material, in other embodiments the access port is molded into the rim wall, and in other embodiments a third vessel surface that is heat sealed to the extension or the rim wall and the flange forms the access port. Upon completion of introduction of the vessel contents, the rim and flange extensions may be folded and heat sealed to seal the conduit, after which the distal portion of the extension beyond the seal may be removed by cutting at the seal boundary.

[0025] In some embodiments, one of the container shell portions includes a flange, a planar wall offset and parallel to the flange, and a continuous swept rim interposed between the planar wall and the flange, where the swept rim includes a curvature that allows the dimension between the flange and the planar surface to be reduced or expanded by some amount. Also, the swept rim profile is continuously widening from the planar surface junction to the flange junction such that solidified contents molded within the interior cavity of the container shell portion are removed in a vector perpendicular to the planar wall without being interfered with by the container shell portion. In some embodiments, the container shell portion described is sealed by thermal bonding (such as heat welding) of the flange rim to the planar portion forming a container having two flat parallel surfaces, one planar surface having a larger area than the other planar surface, and forming a closure on the shell portion.

[0026] In some embodiments, the vessel comprises an interior volume, shape, and features configured such that when the vessel contains a solidified aqueous solution and when a heater surface is placed in contact with the exterior of the larger area planar vessel wall of the vessel, a means for driving the solidified mass into contact with the interior surface of the larger area planar vessel wall results in a rapid phase change of the solid aqueous mass. In some embodiments, a planar sweep or driver card member is provided inside the vessel described. In some embodiments, the driver card includes a thin sheet structure including a material to which a displacement force is applied when the material is placed in a magnetic field, the thin sheet structure being fully laminated within two layers of the material of which the vessel wall is constructed. In some embodiments, the material of the thin sheet structure is a ferritic material. In some embodiments, the ferritic material is a stainless ferritic steel.

[0027] In some embodiments, one or more shell parts include one material lamination. In some embodiments, one or more shell parts include multiple material laminations. In some embodiments, the electronic circuitry is embedded in or interposed between the laminations of layers. In some embodiments, the electronic circuitry carried between the shell laminations may further include electronic sensor circuitry. In some embodiments, data representative of the state of the physical properties of the container contents may be collected by the electronic circuitry carried between the laminated layers of the container shell parts. In some embodiments, the electronic circuitry includes data coupling structures exposed to the external environment, such that external instrumentation or machinery can exchange electronic signals with the circuitry contained between the laminations of the shell parts by contacting such exposed circuitry.

[0028] In some embodiments, the container of the present invention is subdivided into two or more separate chambers. In some embodiments, the two or more separate chambers are interconnected. In some embodiments, the two or more separate chambers are interconnected by a passageway, including a passage, a conduit, a valve, a gate, an opening, a lumen, a tube, a flow passage, a tunnel, a through hole, or other similar compatible structure. In some embodiments, the separated chambers are separated by a swept rim boundary adjacent to a flat conduit of one of the container shell parts, where similar or adjacent regions of the two container shell parts are fused together, for example, by heat sealing. In some embodiments, the container receiving volume is formed by cavities on both shell parts. In some embodiments, the container receiving volume is formed by one or more cavities formed on a single shell part, where the one or more cavities are closed by a planar shell surface on the second shell part or the mating shell part. In some embodiments, the separation of the chambers is interrupted by a conduit connecting the otherwise separated chambers. In some embodiments, the conduit connection is selectively opened or blocked by a gate or valve mechanism to perform a specific function. In some embodiments, the gate or valve mechanism is actuated by an influence external to the sealed shell wall of the container. In some embodiments, the influence of the gate or valve actuation includes an electric field, a magnetic field, a centrifugal force. In some embodiments, two or more chambers are filled with different fluids. In some embodiments, suspended particles may be encouraged to move from one chamber to the next by a centrifugal force, a magnetic field, or an electric field. In some methods, cells may be present in suspension in one fluid in a chamber of the container and then transferred to another fluid in another chamber of the container with minimal mixing of the two fluids. In some methods, cells suspended in cryoprotective medium in one chamber of the container may be transferred under the influence of centrifugal force to another chamber of the container containing recovery medium or injection medium with minimal mixing of the two media. In some methods, the cells are concentrated during the transfer process and in other methods, the cells are diluted during the transfer process.

[0029] Some of the features of the present invention are generally described in FIGS. 1 to 16C, which are provided for the purpose of illustrating some embodiments of the present invention and do not constitute a limitation of its scope of protection.

[0030] 1, an exploded view of a shell storage vessel or shell containment vessel of the present invention is shown. In some embodiments, the containment vessel includes a first / upper / top shell portion 135 and a second / lower / bottom shell portion 180, each including a major flat surface 137 (represented on both portions) and a flange 185, both of which are connected all around by a swept rim 187. The swept rim profile provides additional functionality and is described in further detail in subsequent figures. The two vessel shell portions 135, 180 are joined by thermal welding at the surface of the flanged rim 185 such that the interior of the resulting vessel volume is completely isolated from the exterior on all surfaces and points.

[0031] In some embodiments, the container shell parts include a material that retains toughness, flexibility, and puncture resistance properties at cryogenic and ambient temperatures. In some embodiments, the material is a copolyester polymer. In some embodiments, the copolyester polymer is Tritan™ copolyester, and in other embodiments, the copolyester polymer is Tritan copolyester MP100 available from Eastman Chemical Company. In some embodiments, the shell parts may be formed from sheet material by a process of vacuum forming. In other embodiments, the shell parts may be formed by an injection molding process. In some embodiments, the thermoplastic shell part materials may be joined by heat fusion bonding. In other embodiments, the welding process may be ultrasonic welding, and in other embodiments, the welding process may be laser welding.

[0032] In some embodiments, the flange rim 185 of the container is received within a slot recess 195 (described in more detail in subsequent cross-sectional views) received within a bumper frame portion 190 that may surround some or all of the circumference of the container. In some embodiments, the bumper is installed during any combination of stages of the shell container's use cycle, including pre-filling, filling, freezing, storage, shipping, and thawing stages, to protect the flange joint and rim periphery of the container. In some embodiments, the bumper frame is constructed from an adhesive laminate of felt material, and in other embodiments, the bumper is constructed from a foam material. In some embodiments, the bumper felt material is polypropylene felt. In some embodiments, the laminate of felt material is joined by stitching, and in other embodiments, the laminate of felt material is joined by heat welding or by adhesive bonding.

[0033] In some embodiments, the major planar surface of the top shell 135 includes one or more recessed access ports or port bays 145, 140, 150 molded therein. The port bays have features that are described in more detail in subsequent figures, but generally, each port bay or bay includes a surface that is angled relative to the major planar surface 135, and each angled surface further includes a molded target feature. In some embodiments, the molded target feature indicates a location where the thickness of the container wall is not changed or is reduced in thickness to reduce the force required to penetrate the material with a tubular needle or access device. In other embodiments, the molded target feature includes a recess or tapered conical entrance to facilitate engagement with a matching tapered or threaded adapter. In some embodiments, the orientation of the angled surface of the port bay is controlled to provide a functional orientation to the feature, particularly with respect to the angle of inclination in a gravitational field that facilitates the use of or optimizes the results of the various steps and methods applied for use in the present invention. Thus, in some embodiments, some port bays are assigned a functionality and nomenclature. For example, port 150 is angled and oriented such that the port may be optimally used to fill the container when the corner in which it is located is elevated above the remaining corners of the container, and as such may be referred to as a "fill port." Similarly, port 140 may be elevated above all other corners of the container to provide venting of the container's interior volume when the port is pierced, and may be referred to as a "vent port," while port 145 may be optimally used to withdraw the contents of the container when held at a lower level than the other ports, and as such may be referred to as an "extraction port." While the exemplary embodiment shown in FIG. 1 shows a particular number of port bays in defined locations and is described as having particular functions, illustration is not intended to be limiting, and indeed the number and location of port bays, as well as the intended functional use of any port bay, may vary in other embodiments, and the unlimited configuration options provided by the described port access is one of the primary advantages of the port bay design.In some embodiments, the port bay may be exclusively confined to one of the two shell parts, while in other embodiments, the port bay may be present in both shell parts or may be attached to an additional part that is joined to the two shell parts. In embodiments where the port bay is recessed below the shell planar major surface, the occlusion seal 160, 165, 170 may be non-permanently bonded to the shell surface to isolate and seal the recess of the port bay. In some embodiments, the seal includes an attached tab that is not non-permanently bonded or is weakly bonded to the shell surface such that the tab may be used to remove the seal from the port. In some embodiments, the seal is airtight and can maintain sterility of the port bay following sterilization of the entire container assembly. In some embodiments, the seal is a laminated polymer and metal foil seal.

[0034] Those skilled in the art of using cryogenic storage containers will recognize that all currently available containers are associated with the difficulty of withdrawing the entire contents of the container, often forcing the operator to discard a portion of the contents within the container. In many cases, this difficulty may be imposed in part by the characteristics of the beveled hypodermic needle used to withdraw the contents of the container, as the tip of the beveled needle begins to aspirate the gaseous contents of the container, thereby disrupting the vacuum-induced pressure differential used to withdraw the contents. In some embodiments, the present invention provides a port 155 located at a position on the rim wall, which allows the beveled portion of the needle to contact the corner seam inside the container at the precise acute angle of the needle tube bevel (e.g., typically 12 degrees), thereby facilitating the entire contents of the container to be removed by vacuum suction. In future references, this port may be referred to as the "last drip port." Similar to the ports previously described, the last drip port 155 may be blocked by a seal 175 until specific access to the port is required.

[0035] Since the seals 160, 165, 170, 175 typically comprise thin materials that may be vulnerable to puncture in some embodiments, a protective cover overlay 105 molded to match certain dimensions and contours of the shell container may be placed over the shell surface to protect the port bay seal during storage and shipping operations in some embodiments. In one embodiment, the protective cover 105 is held in contact with the container shell assembly by a flange extension 130 that is attached to the cover 105 along a portion or the entire length of an end of the cover rim 131. In some embodiments, the cover rim flange 130 is received within a slot 195 of the felt bumper 190 that matches with a rim flange 185 of the upper container shell 135. In some embodiments, the absence of the cover flange 130 at the corner 125 of the cover allows the flange 130 to flex and momentarily displace the felt of the felt bumper 190 when an upward force is applied to the cover, thereby allowing the cover 105 to be released from the container and bumper assembly by this means. In some embodiments, the cover 105 is held in place by only two cover flanges 130, while in other embodiments, more than two cover flanges are provided. To facilitate removal of the cover from the shell container and bumper assembly, in some embodiments, a pull tab feature 115 is provided as a partial cutout in the major plane of the cover piece 105. In some embodiments, a slot cutout 110 is provided, which allows the tab to be pulled up, thereby breaking two perforation lines 120 in the material further allowing for pulling of the pull tab 115. Continued pulling on the pull tab 115 may deflect the flange 130 contained within the bumper slot 195, thereby facilitating removal of the cover from the container and bumper assembly.

[0036] 2A, 2B, and 2C, various perspective views of an assembled container assembly are shown to provide greater detail of the invention previously described and illustrated in FIG. 1. FIG. 2A shows the container assembly 200 including the containment vessel 205, bumper 210, and sterile port bay cover 215 of the present invention as it appears fully assembled prior to applying the filling procedure described in the subsequent figures. More detail of the container assembly features may be observed in FIG. 2B, where a cross-sectional side view is shown. In the illustrated embodiment, the upper shell portion 220 is heat welded to the lower shell portion 225 along a rim flange 235, thereby forming a containment volume 230 into which liquid contents may be introduced. In some embodiments, the shell material includes one single material that forms the container container that completely surrounds the interior containment volume 230 of the container and exclusively includes heat welded seams. In other embodiments, the container may include multiple materials where the seams between the different materials are not heat welded prior to the filling and sealing steps, but after the filling and sealing steps, these embodiments may include a heat welded seam that is heat welded to the lower shell portion 225. (a) In contact with the inside of the container, (b) completely encloses the interior containment volume of the vessel 230; and (c) exclusively contains heat welded seams; The protective bumper 236 includes one single layer of material. In some embodiments, the container weld rim flange 235 is captured within a slot recess of the protective bumper 236. In some embodiments, the protective bumper includes two or more layers of impact absorbing material. In the embodiment shown, the middle layer 238 of the bumper 236 is sandwiched between the two outer layers 237 and 240 of the bumper 236, where three layers are bonded together. In some embodiments, the layers are bonded by a suitable bonding method, such as, but not limited to, a stitch bond, a heat fusion bond, or an adhesive bond. In some embodiments, the bumper includes a felt material. In some embodiments, the felt material is polypropylene felt.

[0037] 2C shows the bumper and the interaction between the container and the bumper in more detail, where the top shell portion 245 and the bottom shell portion 250 are heat welded together at the flange rim 256, which also meets at the outer edge with the intermediate bumper laminate layer 276 sandwiched between the upper bumper layer 275 and the lower bumper layer 277, so that an impact with a vector lateral to the outer surface of the bumper is absorbed by the bumper material such that the load is evenly distributed along the outer edge of the container flange rim 265. Additionally, in some embodiments, the bumper frame on the upper lamination 275 and the lower lamination 277 engages with the swept rims 270 of the top shell portion 245 and the bottom shell portion 250, thereby increasing the area along which the compressive load can be distributed, thereby increasing the protective properties of the bumper. In some embodiments, when the protective cover is engaged with the container and bumper assembly, the volume 271 between the bumper and the container flange and swept rim may be occupied by a portion of the protective cover portion (not shown; see 105, 130 in FIG. 1). In some embodiments, the bumper is a continuous frame into which the shell container may be inserted by one distortion of the laminate. In other embodiments, the felt bumper frame is interrupted with lap or scarf joints (not shown) that facilitate installation and removal of the container from the bumper.

[0038] In some embodiments, the swept vessel rim of one or both of the shell top portion 245 or the shell bottom portion 250 includes a curvature such that the distance between the planar surfaces of the upper shell 246 and the lower shell 247 increases or decreases while allowing the surfaces to remain planar through a change in the curvature of the rim profile 270. It will be appreciated by those skilled in the art that aqueous solutions undergoing a solidification phase change will expand, thereby imposing substantial pressures on any vessel containing liquid. A vessel wall that is inflexible will likely rupture under the expansion pressure, while a vessel wall comprising a flexible material will bulge and flex against the expansion pressure. In the embodiment shown in FIG. 2C, distinct performance advantages may be realized if the shell planar surfaces 246, 247 remain planar after the phase change of the vessel contents, and thus applying an external shape to the vessel surface that allows for imposing a final planar configuration on the surface while allowing for an increase in the separation distance between the two surfaces would provide for the realization of these advantages. In some embodiments, the swept rim profile curvature allows for a change in the distance between the upper shell surface 246 and the lower shell surface 247. In some embodiments, the material comprises a material that retains some degree of plasticity over a sufficient aqueous phase change temperature range to allow the necessary change in curvature. In some embodiments, the material is a copolyester polymer. In some embodiments, the copolyester is a Tritan copolyester, and in some embodiments, the Tritan copolyester is MP100 provided by Eastman Chemical Company.

[0039] Referring now to FIG. 3, a dimensional diagram of the embodiment described in FIG. 1 and FIG. 2 is shown. In some embodiments, the container includes an internal volume of about 220 ml. It can be appreciated by those skilled in the art that all dimensions of the container design are independently scalable, and thus containers having a large range of volumetric capacities can be constructed. It can be appreciated that keeping the thickness dimension at or below the approximate values ​​shown provides the advantage of a large surface area to volume ratio, which allows for the characteristics of rapid removal or introduction of thermal energy into the container. In some embodiments, the volumetric capacity of the container will range from 1 ml to 1000 ml. In other embodiments, the volumetric capacity will be greater than 1000 ml or less than 1 ml.

[0040] 4A, a detailed cross section through a corner of a shell vessel is shown where one embodiment of a recessed port bay is installed. In some embodiments, an upper shell portion 405 mates with a lower shell portion 406 and is joined at the interface of a flange 415 with a heat weld 410 bond. A recessed port bay feature 420 is introduced into the upper surface of the upper shell portion 405 during the molding process. The recessed port bay feature 420 includes a flat surface 422 disposed at an acute angle relative to the vessel surface 405 to which the port is attached. In some embodiments, the angle of the flat surface 422 relative to the vessel surface 405 may be other than the angle shown in the drawing. In some embodiments, the orientation of the normal vector to the flat surface 422 relative to an axis perpendicular to the upper surface of the vessel 405 may take any angular value. In some embodiments, a recess feature 425 is molded into the central region of the flat surface 422 and terminates at a lower surface penetration target 430. In some embodiments, the recess feature is a tapered conical section as shown. In other embodiments, the dimple feature may include alternative shapes. In some embodiments, the dimple feature includes a spherical cross section. In some embodiments, the depth dimension of the dimple may be shorter or longer than that shown in the drawings. In some embodiments, the dimple feature may include a radius value that is shorter or longer than that shown. In some embodiments, the dimple feature is not present.

[0041] 4B, one embodiment of the recessed port bay described in FIG. 4A shows a reduced relative thickness of the penetration target 440 of the indentation feature 425. It can be appreciated by those skilled in the art that thermoplastic materials such as copolyester materials have greater toughness and resistance to puncture compared to rubber materials such as those contained in well-known commercially available needle-like septum stoppers. Thus, in some embodiments, it may be necessary to adjust the thickness of the intended target area for penetration of the shell wall by an access instrument such as a hypodermic needle, symmetrical point needle, or bag port spike to modify the force required for penetration. Thus, in some embodiments, following molding of the container shell portion, the indentation target 440 may be modified in thickness by localized heating and molding, and by this method the thickness may be precisely adjusted to the appropriate dimension. In some embodiments, the thickness of the adjusted area is uniform, while in other embodiments, the thickness of the adjusted area varies across the area 440.

[0042] 4C, another embodiment of the recessed port bay described in FIG. 4A is shown in which the recess feature 425 is filled with a different material 445 than that used for the vessel shell. In some embodiments, the material is a rubber material. In some embodiments, the fill material is secured to the recess feature by a thin cap of material 455. In some embodiments, the material of the thin cap is the same as that used for the vessel shell. In some embodiments, the thin cap is heat welded around perimeter 460 to the corner face 465 of the recessed bay port. In some embodiments, the thin cap material and the fill material include a dimple target 456 to facilitate correct placement of the end of a penetration tool.

[0043] 4D, another embodiment of the recessed port bay described in FIG. 4A is shown where recessed port bay 420 is filled with material 475. In some embodiments, the filling material is a rubber material. In some embodiments, target dimples 480 are molded into the top surface of the filling material. In some embodiments, the rubber material is flush with the surface of the container, while in other embodiments, the material only partially fills the recessed port bay.

[0044] 5A-5E, an example method for filling, sealing, and preparing the vessel container described in the previous figures is shown and described. In FIG. 5A, a representative vessel fill port corner 500 of a vessel is shown oriented such that the fill port corner is at a higher elevation than the remainder of the vessel. In the described example, the vessel container shell 507 is surrounded by a protective bumper 505 around the flange, however, in some vessel filling protocols, the bumper 505 may be absent and may optionally be installed in post vessel filling operations. In some filling protocols, the filling operation may be performed in a clean room or sterile area isolated by a barrier that maintains the sterility of the local filling environment. Covering the fill port is a label seal 510 that is reversibly adhered with an adhesive to the surface of the vessel shell 507, thereby isolating the fill port recess and maintaining the sterility of the port recess. The label seal 510 may be removed by applying an upward tension on the removal tab 511, thereby initiating separation of the label from the surface of the vessel shell 507. In some embodiments, the removal tab 511 is not adhered to the container surface 507, thereby facilitating lifting of the tab. In some embodiments, the removal tab 511 may include an extension or a through hole to facilitate gripping by mechanical filling machinery.

[0045] 5B, when the label seal 510 is removed, the fill port recess 515 is exposed, exposing the access target 520. In some embodiments, the access target is a simple depression in the wall of the recess. In some embodiments, the access target takes on a more complex profile, such as a truncated conical depression, and in other embodiments, the access target includes more complex features, such as, but not limited to, a twist lock engagement structure or a threaded connection. In some embodiments, the access target further comprises a material other than the material from which the container wall 507 is constructed.

[0046] 5C, the access target is pierced by a fill needle 525, which in some methods may be driven by a mechanical filler element 520. In some methods, the fill needle 525 has dual counterflow passages, i.e., one passage through which liquid may be introduced into the container and another passage through which gas from the container interior may be withdrawn, thereby maintaining pressure equilibrium between the container interior and the atmospheric environment in which the container is resting. When the container is filled to the desired containment volume, in some methods, the fill needle 525 is withdrawn.

[0047] 5D, in some methods, after filling, the access target is sealed by applying a sealing element 535 to the access target 520 of the fill port recess 515. In some embodiments, the sealing element is a plug shape. In other embodiments, the sealing element is a cover, and in other embodiments, the sealing element is a combination of a cover and a plug shape. In other embodiments, the sealing element may include a cup shape to function in subsequent uses such as venting or extracting the container contents. In some embodiments, the material of the sealing element is the same as that of which the container is constructed. In some methods, following application or insertion into the access target 520, the sealing element is heat impulse sealed (sealing mechanism not shown) to the surface of the fill port recess 520 such that fusion of the material occurs, thereby completely sealing the container interior from the outside environment.

[0048] 5E, in some filling methods, the final step is to apply a sterile seal cover 545 to the container shell surface 507 such that the fill port recess (515 in the previous figure, here blocked) is isolated from the outside environment. In some embodiments, the seal cover 545 does not include a lifting tab (515 in the previous figure), as shown in this figure, to specify to the user that this port will not participate in subsequent use operations such as venting or withdrawal of the container contents. In other embodiments, a tab feature (515 in the previous figure) may be present, thereby facilitating removal of the seal 545 for subsequent use operations.

[0049] Now referring to FIG. 6A, an example of a sterile venting tool is shown in perspective and cross-sectional views. In some embodiments of the present invention, gas is vented from the external environment into the container interior to replace the volume of liquid displaced during extraction of the liquid contents. In some embodiments, sterility within the container interior is maintained during the liquid extraction process by passing the external gas through a filter with a maximum pore size sufficient to retain pathogenic organisms and prevent their ingress into the container interior. In the illustrated example 600, a cylindrical container housing 610 is shown that includes a cylindrical interior volume that is open at the top so that the interior volume is continuous with the external environment. Housed within and occupying the entire space of the cylindrical interior volume is a filtration element 605 that has a maximum pore size such that any pathogenic biological material is trapped within the filtration material in addition to any particulate matter while allowing gas to pass through the filtration medium. Filtered gas passing through filter 605 passes under plenum region 612 and is directed into hollow portion 613 of tube 615 contained within cylindrical housing base 614 and out tube end 616 (shown here as a sharp bevel in tube 615). In some embodiments, the filter (as a built-in accessory) may be attached to a needle cover that reversibly bonds by friction to a cylindrical extension 617 of the filter housing, thereby maintaining sterility of the needle until use.

[0050] Referring now to FIG. 6B, a method for venting an embodiment of a container of the present invention is shown. In the illustration, a cross section of a vent port corner of a container shell 650 and a bumper felt bumper frame 655 is provided. A sterile vent tool 610 (as described in FIG. 6A) is shown inserted through the rubber filler 630 filling the vent port 640, which is continuous with the container top shell 650, and through the access target 635 of the recessed port 640, thereby penetrating the continuous shell boundary that previously completely surrounded the container interior 645, and thereby forming a path through which gas from the external environment may pass to the container interior 645 by flow through the sterile filter (element 605 in FIG. 6A). In some embodiments, the access target material 635 at the boundary of the filtration vent needle 615 forms an airtight seal sufficient to prevent gas flow between the access target material 635 and the filtration vent needle 615. In other embodiments, an airtight seal is formed between the filler 630 and the filtration vent needle 615, and in other embodiments, the seal with the filtration vent needle is formed by other structures, such as, but not limited to, the filler 445 and cap structure 455 shown in FIG. 4C.

[0051] 7, a cross-sectional view of an embodiment of the present invention is shown in which a first shell portion 705 is mated to a second shell portion 710 at a flange interface 712 of both portions, with a port recess 715 present in a major surface 717 of the first shell portion 705. Also, a through hole is introduced in one surface 730 of the port recess 715, into which a threaded insert portion 720 is inserted such that an underside of a flange 725 of the insert 720 contacts and fits against the surface 730 of the port recess 715. In some embodiments, the threaded insert 720 is heat welded to the port recess surface 730 such that the interior of the vessel 740 is isolated from the environment outside the vessel. In some embodiments, the insert 720 is constructed from the same material used to form the first shell portion 705 and the second shell portion 710. In some embodiments, the insert 720 is made by an injection molding process. In some embodiments, the insert 720 includes internal threads 745 that allow an external coupling device to positively interact with the recess port insert. In some embodiments, one lower wall of the insert 720 may include a second thickness 735 to adjust the force required to penetrate the wall by an instrument when coupled to the insert 720.

[0052] 8A, an embodiment of the present invention is shown in cross section. In this embodiment, an upper shell portion 805 is joined to a second or lower container planar shell portion 810 in a fusion bond between the material around the peripheral flange 820 of the upper shell portion 805 and the opposing material of the second or lower shell portion 815. Those skilled in the art will recognize that when filled with an aqueous solution and placed in an environment with a temperature below the freezing point of the solution, expansion of the contents will occur during the freezing process. In some embodiments, to allow for expansion of the container under the outward pressure of a material undergoing a phase change on the interior surface of the container, the swept rim 825 may include a profile with a curvature 835 such that the change in curvature when pressure is applied to the inner surface of the profile allows the distance between the two shell portion surfaces 825, 810 to be increased without distorting the shell portion planar surfaces 805, 810 and without overstressing the fusion bond 830 between the upper shell rim flange 820 and the opposing portion 815 of the second shell portion 810. In some embodiments, an external shape may be applied to the container during the freezing process to limit shape changes to the container shell wall other than the swept rim 825.

[0053] 8B, a cross-section of the container shown in FIG 8A is shown in which swept rim 825 includes a more exaggerated double back curvature 835 than its rim counterpart shown in FIG 8A. In some embodiments, the length of the profile line defining swept rim 825 increases as the distance between upper shell portion 805 and lower planar shell portion 810 increases to allow for a greater range of extension of the swept rim.

[0054] 9A, an embodiment of the invention includes a shell of a first or upper shell portion 905 including a major planar surface 930 with rounded corners joined with an expandable swept rim 927 further joined with a flange rim 910, which is joined with a planar second or lower shell portion 965 (closed behind the upper shell portion 905). Intercepting the flange rim of the first shell portion 905 is an extension 920 of the swept rim 927 which forms all sides of a conduit tunnel except for the side formed by the second portion, which forms a passage between the vessel interior volume and the outside environment. In some embodiments, the conduit tunnel terminates in an end plug 945 heat welded to the material forming all sides of the conduit tunnel, which further includes one or more through holes, openings, lumens, or passages through which material may be introduced into the vessel interior. In some embodiments, the through hole in the end plug 945 is closed by a reversibly bonded airtight seal label 950 configured to isolate the interior of the container until removed. In some embodiments, the vent port 955 and extraction port 960 are located within the wall of the swept inflatable rim 927. In some embodiments, the flange 910 is joined with an extension 935 that can function as a handle for manually or mechanically moving the container, thereby isolating the remainder of the container from the influx of thermal energy from the gripping element. In some embodiments, the surface flange extension 935 provides a flat surface onto which an identification label 940 can be affixed. In some embodiments, the flange extension 935 and affixed label 940 allow the container 900 to be stored in a horizontally stacked configuration with the extension and label extended upward to facilitate rapid identification and location of a selected container from a collection of similar containers. In some embodiments, flange extension 935 and attached label 940 serve to reduce the time required to locate a particular container, thereby mitigating the temperature rise and damage to contained samples associated with keeping a collection of containers at room temperature while such a search is performed.In some embodiments, the first or upper shell portion 905 and the second or lower shell portion 965 are partially joined at the flange extension 935 such that a double-sided container is formed between the two portions. In some embodiments, the flange-extended double-sided container may enclose an electronic identification device. In some embodiments, the electronic identification device is an RFID chip.

[0055] 9B, an exploded view of the container described in FIG. 9A is shown to better illustrate the container's construction. In some embodiments, a first or upper container shell portion 905 mates with a second or lower planar container shell portion 965. In some embodiments, the second container shell portion 965 is non-planar and includes a cavity. In some embodiments, the second container shell portion 905 is a nominal mirror image of the first container shell portion at the flange face mating surface. In some embodiments, an injection molded insert plug 945 is heat welded to form a complete seal with the first shell portion 905 and the second shell portion 965 at the through tunnel, through hole, opening, lumen or other passageway formed by the swept rim extension 920 of the first shell portion 905 and the facing region of the second shell portion 965. In some embodiments, the plug cap 945 is reversibly sealed by the attachment of an airtight label cover 950. In some embodiments, the identification label 940 may be affixed to an exterior surface of either the first shell portion or the second shell portion.

[0056] 9C, a first step in the method of filling the container 900 described in FIGS. 9A and 9B is shown. In some methods, the sterile label seal (FIG. 9A, 950) is removed to expose the through passage 922 extending from the top exterior surface of the end cap 945 of the through tunnel 920 into the container interior, and the container 900 is positioned in an upright position with the end cap 945 in the uppermost position. In some embodiments, the liquid contents of the container may be introduced into the container interior by a tubular filling needle. In some methods, an air space remains after introduction of the container contents such that the liquid in the container does not occupy the through tunnel passage 920.

[0057] 9D, a second stage of the method described in FIG. 9C is shown. In some methods, with the container held in an upright position as in FIG. 9C, the filled container penetration tube 920 is collapsed by clamping the passageway 920 with a heating device (not shown) and raising the temperature of the passageway material to the fusion temperature of the glass and material, thereby flattening the passageway to the thickness of the adjacent flange. In some methods, the clamping and heating device includes a trim feature whereby the end cap 945 and adjacent container material may be removed from the remainder of the container at the indicated line 924 such that the trim line coincides with the outer edge of the rim flange at 924. In some methods, the heat compression and heating device seals the container along the path of the container flange such that the flange seal is continuous around the container, thereby isolating the contents of the container from the outside environment.

[0058] 9E, there is shown a third stage of the method described in Figures 9C and 9B. In some methods, once the sealed and trimmed container along the flange perimeter 932 has cooled, the sealing of the container is complete and the container and contents may be further processed by subsequent methods, such as freezing, storage, shipping, or thawing methods.

[0059] Referring now to FIG. 9F, a dimensional drawing of the embodiment shown in FIGS. 9A-9E is shown. The container embodiment shown has a nominal volume capacity of 2 ml. In some storage methods, the dimensional drawing shown may be stored in a typical 5 inch by 5 inch cryogenic storage box with 3 columns and 27 rows to accommodate a total of 81 containers per box. In some methods, without prior knowledge of the container location, the identity of a particular container may be visually assessed by scanning the rim extension label without handling the container and lifting it, as would be required for a similar array of cylindrical tubular vials with labels affixed to the side.

[0060] 10A, an exploded view of another embodiment of the present invention is shown showing a first or upper shell container portion 1005 having an interior volume boundary consisting of a flat planar surface 1015 bounded by a complex profile swept rim 1016 and a peripheral flange 1022 extending from the lower end of the rim 1016, in which one or more recessed port structures 1017 are housed. In some embodiments, the second or lower shell container portion of the container is a flat planar surface portion 1010 that mates with the upper shell container portion 1005 where the flange rim 1022 meets the peripheral rim 1020 and is joined by a heat weld or other suitable joint, thereby forming a container shell body having two parallel opposing flat planar surfaces. In some embodiments, the swept rim 1016 flares outwardly from the top surface 1015 without an undercut surface such that contents that solidify with the container upon release from the top shell inner surface are permitted to move in a directional vector perpendicular to the top plane of the container without interference from the top shell container portion 1005. In some embodiments, a driver card element, component or part 1025 is contained within the vessel interior volume. In some embodiments, the driver card 1025 includes a ferrite plate 1035 laminated between two layers of material 1030 that are joined at the edges with a heat weld or other suitable bond such that the ferrite plate 1035 is completely surrounded by the laminate material 1030. In some embodiments, the laminate material 1030 is the same material used for the vessel shell parts.

[0061] 10B, the top shell 1015 is shown joined to the lower planar surface 1010 with a heat seal bond 1040 to form the complete container structure. In some embodiments, the lower planar surface 1010 allows a planar heater surface to contact the lower surface, thereby increasing the temperature of the contained contents.

[0062] 11A, a first stage of a thaw sequence progression is shown to illustrate how freeze-thawing of the container contents may be performed. In FIG. 11A, a cross-sectional view is shown of the container top shell 1105 and container planar bottom 1110 described in FIG. 10 being heat welded together around a peripheral flange 1112. Note that the interior volume of the container 1115 for purposes of illustrating the process is filled with an aqueous liquid. The container interior also holds a driver card 1140 including a ferrite plate 1135 laminated between two sheets of the same material used for the container walls (the laminated sheets are shown transparent), the ferrite plate 1135 of the driver card 1140 being held or selectively suspended in contact with the upper inner surface of the top shell portion 1105 via the attractive force of a magnet 1130 embedded in a block of thermally conductive material 1125. An additional block 1120 constructed of thermally conductive material is in direct contact with the lower surface of the container 1110. At this stage of the method, both blocks 1125 and 1120 are lowered in temperature such that thermal energy is conducted away from the container and container contents until a phase change occurs in the liquid contents. Upon solidification, the top block 1125 and magnets 1130 and bottom block 1120 are removed, leaving the driver plate 1140 trapped in a stationary position by the solidified contents.

[0063] 11B, the vessel and contents 1105 described in FIG. 11A are shown in a solidified state. At this step in the method, the upper shell 1105 and the lower planar shell portion 1110 forming the complete vessel are now clamped between a heater block 1150 in direct contact with the upper planar surface of the vessel 1105 and a second heater block 1155 in direct contact with the lower planar surface of the vessel 1110. The lower heater block 1155 has magnets 1160 embedded in it. When the temperature rises to a value sufficient to cause the solidified contents of the vessel portion 1105 to separate from the inner surface of the upper shell, the magnetic attraction of the magnets 1160 on the driver card 1040 biases the solidified contents against the inner surface of the lower flat vessel portion 1110. At this point in the method, the upper heater block may be de-energized or alternatively pulled away from contact with the outer surface of the upper vessel shell portion 1105 so that it is no longer in a heating mode. As the phase change of the solid container contents 1105 occurs at the inner surface of the planar container shell portion 1110, the bias pressure of the driver card 1140 forces the liquid formed at the inner surface of the portion 1110 to the periphery and then accumulates between the driver card 1140 and the inner surface of the upper shell portion 1150. As this process progresses, only the solidified container contents 1115 will be in contact with the inner surface of the flat container portion 1110, thus acting as an efficient heat sink for the thermal energy provided by the lower heater block 1155; the temperature at the solid material 1155 remains close to the phase change temperature of the solid material 1115; therefore, a high wattage influx of thermal energy may be applied to the thawing process resulting in a reduction in the phase change interval without the risk of damage to the liquid contents due to temperature increase.

[0064] 11C, the defrosting process is shown at a later time than in FIG. 11B. At this stage of the defrosting process, the upper heater block 1150 is shown still in contact with the top flat surface of the upper shell portion 1105, but is de-energized. As the defrosting process progresses at this stage, the solid material contents 1105 in the container are reduced and the driver card 1140 is in a lower position in the container. Similarly, the volume of liquid contents accumulated in the space 1175 above the driver card 1140 is increased. During this stage of the defrosting process, the heater block 1105 may still be generating a high wattage output for heat energy in some embodiments. In some embodiments, the upper heater block 1010 may be actively cooled to control the temperature of the liquid contents in the space below it. In some embodiments, the proximity of the driver card may be monitored by a Hall sensor, or an inductive sensor (not shown), embedded in or near the heater block 1105 to determine the state of the defrosting process and to determine the ultimate completion of the defrosting process.

[0065] 11D, there is shown the result of the thawing process described with reference to FIGS. 11A-11C. At this stage of the thawing process, the upper heater block 1150 remains de-energized and is in contact with the top flat surface of the upper shell portion 1105. The solid contents 1115 inside the container have completed a phase change to a liquid phase and are displaced into the space 1175 above the driver card 1040, which is now biased against the inner surface of the lower flat container portion 1110 by the magnet 1160. At or before this point in the thawing process, the heater block 11055 has been de-energized and is no longer transferring heat to the container interior. In some embodiments, the heater block 1155 and / or heater block 1150 may be actively cooled to maintain a desired operating temperature for the liquid contents of the container following the thawing process.

[0066] 12, an exploded view of an embodiment of the present invention is shown in which a first or upper vessel shell portion 1204 includes a planar surface 1205 joined at a peripheral edge to a swept rim 1210 which is further joined at an opposite end by a flange 1215 occupying a plane parallel to and offset from the planar surface 1205. When assembled, the flange 1215 mates with a matching area of ​​a second or lower planar shell piece 1235 and is heat welded to form a containment vessel that completely isolates the interior volume from the environment. In some embodiments, the driver card 1220, including the ferritic sheet material 1230 laminated between two layers of material 1225 such that the ferritic sheet material 1230 is completely surrounded by the material 1225, thereby isolating it within a watertight containment vessel, is enclosed within the containment vessel formed when the top shell portion 1204 is joined to the bottom shell portion 1235. In some embodiments, the lower shell portion 1235 is longer than the upper shell portion 1204 such that when the upper shell portion 1204 is joined to the lower shell portion 1235 with the edges 1206, 1236 mated, the opposite end of the container shell portion 1245 overhangs and extends beyond the upper container shell edge 1216. In some embodiments, the lower shell portion 1235 comprises two heat-welded laminates of material. In some embodiments, the two laminates 1235 comprise a sensor 1250 interposed between the laminates and connected by a conductive material trace 1255 that terminates on the extended edge 1240 of the lower shell portion 1235. In some embodiments, the sensor 1250 is a temperature sensor. In some embodiments, the sensor 1250 is a thermistor, RTD sensor, or thermocouple sensor. In some embodiments, the sensor communicates directly with the container interior or container contents through an area where there is no internal laminate, and in other embodiments, a material other than the container shell material is interposed between the sensor and the container interior or container contents. In some embodiments, a portion of the top laminate layer 1238 is cut away to form 1239 exposing an end portion of the conductive sensor trace 1240 .In some embodiments, the conductive sensor imprint can be interfaced with a complementary electrical socket so that an electronic data signal from the sensor 1250 can be communicated to data processing circuitry. In some embodiments, the top layer 1238 of the lower shell portion 1235 is thinner than the second layer such that the temperature registered by the sensor 1250 correlates closely with the top surface of the top layer 1238, and thus with the temperature of the liquid contents contained within the container.

[0067] 13A, an embodiment of the present invention is shown, where a container 1300 includes an upper shell portion further including two planar surfaces, a first planar surface 1310 contiguous with a swept rim structure 1335 that forms a side of a first chamber 1311 of the upper shell portion, and a second planar surface 1315 that contiguous with a swept rim structure 1340 that forms a side of a second chamber 1316 of the upper shell portion. Both swept rim structures 1335, 1340 are contiguous with a rim flange 1305 that surrounds both chambers 1311 and 1316 and is contiguous with a flange 1306 that partially divides the two chambers, and both swept rim structures 1335 and 1340 are contiguous with a rim flange 1307 that further divides the two chambers 1311 and 1316. In some embodiments, the two chambers 1311, 1316 are joined by a gas valve 1355 and a valve gate 1350, both of which are normally closed and open upon receiving centrifugal forces aligned with a particular centrifugal field vector running perpendicularly from the proximal end 1312 of the flange rim 1305 to the distal end 1313 of the flange rim 1305. Some embodiments include a loading port 1320 located in the planar surface 1310 of the first chamber 1311. Some embodiments include a loading port 1325 located in the planar surface 1315 of the second chamber 1316. In some embodiments, an extraction port 1330 is located in the planar surface 1315 of the second chamber 1316. In some embodiments, additional vent ports may be located in the planar surface 1315 of the second chamber 1316. In some embodiments, the flange rim 1305 may include two or more through holes or keyhole cutout areas 1345.

[0068] 13B, an exploded view of the embodiment of the container described in FIG. 13A is shown. In some embodiments, a top shell portion 1310 including a peripheral flange 1364 continuous with a flat surface 1368 and a second lower shell portion 1312 including a peripheral flange 1365 continuous with a flat surface 1369 are fitted together with a heat seal that seals the periphery of the container and partially divides two chambers formed by a first chamber volume 1370 of the first shell portion 1310 and a first chamber volume 1371 of the second lower shell portion 1312. Also, a second chamber volume 1373 of the first shell portion 1310 and a second chamber volume 1375 of the second lower shell portion 1312 are joined. In some embodiments, the second surface 1366 of the first upper shell portion 1310 and the second surface 1367 of the second lower shell portion 1312 are thermally welded together to additionally and partially divide the first chamber volumes 1371, 1370 from the second chamber volumes 1375, 1373 such that the first and second chambers of the resulting vessel are separated except where connected by gas valve 1385 and gate valve 1380.

[0069] 14A, an exemplary embodiment of a gas valve that may be used with some of the embodiments described with reference to FIG. 13A and FIG. 13B is shown. In some embodiments, the valve housing 1405 of the gas valve 1400 comprises six exterior flat surfaces 1407 that, in some embodiments, are heat welded to opposing inner surfaces within a container in which the gas valve may be installed. In some embodiments, the valve housing 1405 includes a raised rib structure 1410 that may provide additional material to fill the weld between the surfaces 1407 and the opposing inner surfaces of the container in which the gas valve may be installed. In some embodiments, a septum structure 1415 is held within the valve housing 1405. In some embodiments, the septum structure 1415 is a monolithic molding. In some embodiments, the septum portion 1415 includes a rubber material. In some embodiments, the septum portion 1415 includes a silicone rubber material. In some embodiments, the septum structure 1415 includes an assembly handle 1420 to facilitate insertion of the septum structure into the valve housing 1405. In some embodiments, the septum structure includes a pressure equalization port 1450 and a gas passage port 1431, both of which are described in more detail in subsequent figures.

[0070] 14B, the gas valve described in FIG. 14A is shown in cross section. In some embodiments, a valve housing 1430 having a hexagonal outer surface includes a flexible rubber septum structure 1452. The septum structure 1452 includes a first wall 1455 that communicates with the first chamber of the container (1371 and 1370 in FIG. 13B) and a second wall 1453 that communicates with the second chamber of the container (1375 and 1373 in FIG. 13B). The first wall 1455 includes a pressure equalization port 1450 that, in some embodiments, maintains the pressure in the non-sealing chamber 1451 of the valve at the same pressure as the pressure in the first chamber of the container. In some embodiments, the first wall 1455 also includes a passage port 1431 that is in communication with the valve chamber 1432. In some embodiments, the first wall 1455 seats within the valve housing and forms an annular seal at interface 1440. In some embodiments, the second wall 1453 includes a passage port 1460 in communication with the valve chamber 1432. In some embodiments, the second wall 1453 seats within the valve housing and forms an annular seal at interface 1445. In some embodiments, the valve chamber 1432 is bisected and blocked by the valve gate 1470. In some embodiments, when a centrifugal force is applied along vector 1465, the valve gate 1470 displaces along the same vector 1465, deflecting the adjacent chamber membrane 1480 such that a gap is formed between the valve gate 1470 and the valve housing 1430 at junction 1433. In some embodiments, a dense material 1485 may be embedded in the valve gate 1470 to tune the valve's response to centrifugal force values. In some embodiments, the distance 1454 between the valve gate distal surface 1472 and the valve housing inner surface 1475 may be adjusted to limit the movement of the valve gate 1470 when displaced under centrifugal force load. In some embodiments, the septum structure 1452 comprises a rubber material. In some embodiments, the rubber material is silicone rubber. In some embodiments, an assembly handle 1420 is molded into the septum structure 1452 to facilitate insertion and seating of the septum structure into the valve housing 1430.

[0071] 15A, an exemplary embodiment of a gate valve that may be used in some embodiments described with reference to FIG. 13A and FIG. 13B is shown. In some embodiments, the valve housing 1505 of the gate valve 1500 comprises six exterior flat surfaces 1507 that, in some embodiments, are heat welded to opposing inner surfaces within a vessel in which a gas valve may be installed. In some embodiments, the gate valve housing 1505 includes a raised rib structure 1510 that may provide additional material to fill the weld between the surfaces 1507 and the opposing inner surfaces of the vessel in which the gas valve may be installed. In some embodiments, the gate valve includes a gate housing valve seat 1515, which, in some embodiments, is joined to a V-block structure 1520 via two connector plates 1517, 1518. In some embodiments, a flexible gate 1530 is held within the structure formed by the valve seat 1515, the connector plates 1517, 1518, and the V-block structure 1520. In some embodiments, the top connector plate 1517 includes an opening 1525 through which a flexible gate 1530 can be inserted into the valve structure.

[0072] 15B, a cross-sectional view of the gate valve 1500 described in FIG. 15A is shown. Here, the communication tunnel 1545, which in some embodiments connects the first chamber opening 1540 and the second chamber opening at the valve seat 1560, is seen in cross-section. In some embodiments, permanent magnets 1550 adjacent either side of the communication tunnel 1545 may be embedded in the valve housing 1535. In some embodiments, a flexible gate 1530 is interposed between the valve seat 1560 and the leading edge 1580 of the V-block structure 1520. In some embodiments, the flexible gate 1530 includes two ferrite pieces 1555 that may be attracted by the permanent magnets 1550, thereby holding the closing side 1538 of the flexible gate 1530 in contact with the valve seat 1560, thereby closing the communication tunnel 1545. In some embodiments, when a centrifugal force aligned with vector 1570 is applied to the flexible gate 1530, the closing side 1538 is displaced towards the V-block and out of the way of the valve seat 1560, thereby allowing passage from the first chamber side 1540 of the communication tunnel 1545 to the valve seat 1560 side of the communication tunnel. In some embodiments, when the value of the centrifugal force aligned with vector 1570 is returned to zero, the magnetic attraction of the magnet 1550 on the ferrite piece 1555 causes the closing side 1538 of the valve gate 1530 to again settle onto the valve seat 1560, returning to closing the communication tunnel 1545. In some embodiments, all metallic and magnetic materials are isolated from direct contact with fluids that may flow through any portion of the gate valve 1530 or accumulate around the gate valve 1530. Although the described embodiments have been described as including detailed and specific examples of gas valves and gate valves, it will be understood by those skilled in the art that numerous valve designs may perform the same functions, and thus the examples provided are not intended to be limiting or exclusive to the valve designs shown or the means by which the valves are actuated. For example, in some embodiments, one or more valves may be actuated by a magnetic field, while in other embodiments, one or more valves may be actuated by an electric field.

[0073] 16A, a container embodiment 1608 (shown and described as container 1300 in FIGS. 13A and 13B) is shown attached to a rear half of a retainer 1610 that includes a cavity (shown in FIG. 16B below) configured to receive and contact the exterior of the container 1608 along a major planar surface (closed) and a container rim 1609. Four keyhole slots 1614 of the container 1608 fit over four pin objects 1615 affixed within the retainer 1610, thereby preventing displacement of the container embodiment along the radial force vector 1640. In some embodiments, the lower retainer portion 1610 is secured to a centrifuge arm 1625 by one or more fasteners 1611, such as threaded bolts or screws. The container may be further fully enclosed by affixing four holes (not shown) in the upper retainer 1605 over corresponding pins 1615, which may then be clamped to a centrifuge arm 1625 at two lateral recessed surfaces 1620. Rotation of the centrifuge arm 1625 on its axle 1630 applies centrifugal force to the container 1608 and any contents therein.

[0074] 16B, a cross-section of the assembly of FIG. 16A is shown. In some embodiments, the exterior surfaces of the upper and lower vessel shell portions 1660, 1655 of the vessel 1645 fit closely with corresponding portions of the interior cavity surfaces of the upper and lower retainers 1605, 1610. In some embodiments, the vessel 1645 is subjected to a centrifugal force, and the vessel fluid contents exert an outward hydrostatic pressure on the upper and lower vessel shell portions 1660, 1655, which may distort and stress the vessel, albeit due to the resisting effects of the upper and lower retainers 1605, 1610.

[0075] 16C, the embodiment of the vessel described in FIGS. 16A and 16B is shown with the upper shell portion removed. In some embodiments, the lower vessel shell portion 1655 includes a first chamber 1665 and a second chamber 1666 partially isolated by an intermediate wall 1685 that separates the two chambers along a path. The separation wall 1685 is blocked by a centrifugally actuated valve gate 1675 and a centrifugally actuated gas valve 1656. In some methods, an exchange of fluid containing a large number of cells suspended in the first chamber 1665 may be performed prior to the recovery of the cells from the vessel. In some methods, the first chamber 1665 of the vessel is occupied by a cell suspension that includes a cryoprotectant fluid and a small volumetric percentage of gas, and the second chamber 1666 is occupied by an exchange fluid and a small volumetric percentage of gas. In some embodiments, the proportional volumes of fluid in the two chambers are adjusted such that the hydrostatic pressure difference across the valve gate 1675 is minimized when the container is subjected to centrifugal force along vector 1640 and the contents of both chambers 1665, 1666 are biased to the distal side 1691 of the container. In some methods, the density of the fluid in the second chamber 1666 is equal to or slightly greater than the density of the fluid in the first chamber 1665. In some methods, when the container 1600 is subjected to centrifugal force along vector 1640, the cells in the first chamber 1665 move through a passage tunnel (not shown) of the gate valve 1675 to the second chamber 1666. In some methods, when sufficient centrifugal force is applied, the gas valve 1656 opens, allowing the gas pressure to equalize between the two chambers 1665, 1666. In some methods, when sufficient centrifugal force is applied, the valve gate 1675 opens, allowing cells in the first chamber 1665 to move through the valve gate 1675 and into the second chamber 1666. In some methods, a hydrostatic equilibrium between the fluids in the two chambers 1665, 1666 is achieved, thereby negating any hydrostatic pressure induced displacement of the boundary between the two fluids.In some methods, when a predetermined time has elapsed and / or when it is otherwise determined that the cells in the first chamber 1665 have completely transitioned into the second chamber 1666, the angular velocity of the centrifuge is reduced to zero while the gas valve 1656 and the gate valve 1675 return to their closed positions, thereby again isolating the two chambers. Following centrifugation of the vessel 1600, in some embodiments, the vessel 1600 is removed from the centrifuge and the chamber 1666 is vented at a fill port (not shown), such that the cell suspension in the second chamber 1666 is collected via an extract port (not shown).

[0076] The above discussion of the invention has been presented for purposes of illustration and description. The foregoing is not intended to limit the invention to the form or forms disclosed herein. The description of the invention includes a description of one embodiment and certain variations and modifications, but other variations and modifications are within the scope of the invention, for example, as may be within the skill and knowledge of one of ordinary skill in the art upon understanding the present disclosure. It is intended to acquire rights to include alternative embodiments to the extent permitted, including alternative, interchangeable and / or equivalent structures, functions, scopes or steps to those disclosed herein, and to acquire rights similar to those disclosed herein, without intending to dedicate any patentable subject matter to the public.

Claims

1. A cryogenic storage container for storing a frozen cell suspension, comprising: A first shell portion; and A second shell portion; and a cryogenic storage vessel having an interior volume, the first shell portion has a first major surface, a first inner surface facing away from the first major surface, a first rim, and a first flange, the first major surface including a first length, a first width, and rounded corners, the first rim extending around a periphery of the first major surface between the first major surface and the first flange, the first flange being parallel to the first major surface and extending outwardly from the first rim, and a distance from the first major surface to the first flange defining a depth of the first shell portion; the second shell portion has a second major surface, a second inner surface facing away from the second major surface, a second rim, and a second flange, the second major surface including a second length, a second width, and rounded corners, the second rim extending around a periphery of the second major surface between the second major surface and the second flange, the second flange being parallel to the second major surface and extending outwardly from the second rim, and a distance from the second major surface to the second flange defining a depth of the second shell portion; the first flange and the second flange are joined, and the interior volume is defined by a space between an inner surface of the first shell portion and an inner surface of the second shell portion, wherein the interior volume is isolated from an exterior of the cryogenic storage vessel; and the first rim includes a curved surface having a curvature configured to allow an increase or decrease in a distance between the first major surface of the first shell portion and the second major surface of the second shell portion during a freezing process applied to a cryogenic storage container, allowing the first major surface and the second major surface to remain planar throughout the increase or decrease in the distance between the first major surface of the first shell portion and the second major surface of the second shell portion. Cryogenic storage vessels for storing frozen cell suspensions.

2. the first shell portion and the second shell portion are constructed from a thermoplastic material.

2. The cryogenic storage vessel of claim 1.

3. The thermoplastic material is a copolyester. The cryogenic storage vessel of claim 2.

4. Further comprising a recess formed in at least one of the first and second major surfaces.

2. The cryogenic storage vessel of claim 1.

5. the recess comprises a single annular wall, the annular wall being symmetrical about a central axis of the annular wall; The cryogenic storage vessel of claim 4.

6. The recess includes at least two walls. The cryogenic storage vessel of claim 4.

7. the at least two walls include a partial region having a wall thickness less than a wall thickness of the remainder of the at least two walls; The cryogenic storage vessel of claim 6.

8. the recess is located adjacent at least one rounded corner of the first and second major surfaces. A cryogenic storage vessel according to any one of claims 4 to 7.

9. The cryogenic storage container further comprises: a through hole penetrating at least one of the first and second main surfaces in the recess; an insert within the through-hole for selectively sealing the through-hole to isolate it from the exterior of the cryogenic storage vessel; having the insert is sealed in the recess in at least one of the first and second major surfaces; A cryogenic storage vessel according to any one of claims 4 to 8.

10. the recess is located in the rim of the cryogenic storage vessel; A cryogenic storage vessel according to any one of claims 4 to 9.

11. and further comprising an airtight label removably adhered to the exterior of the cryogenic storage vessel and disposed on top of the well to isolate the volume of the well from the exterior of the cryogenic storage vessel. A cryogenic storage vessel according to any one of claims 4 to 10.

12. a ratio of the first length to the sum of the first width and the first depth is from 1:1 to 4:1; 2. The cryogenic storage vessel of claim 1.

13. a ratio of the first length to the first depth is from 1:1 to 30:1; 2. The cryogenic storage vessel of claim 1.

14. a ratio of the first length to the first depth is from 2:1 to 20:1; 2. The cryogenic storage vessel of claim 1.

15. The area obtained by multiplying the first length and the first width is 3 cm ２ From 300cm ２ That is, 2. The cryogenic storage vessel of claim 1.

16. The area obtained by multiplying the first length and the first width is 300 cm ２ From 1000cm ２ That is, 2. The cryogenic storage vessel of claim 1.

17. At least one of the first and second flanges has a width greater than 1 mm.

2. The cryogenic storage vessel of claim 1.

18. a protective bumper in contact with a peripheral edge of the first and second flanges; 2. The cryogenic storage vessel of claim 1.

19. the protective bumper envelops a portion of the outer peripheral edges of the first and second flanges; 20. The cryogenic storage vessel of claim 18.

20. The protective bumper is made of a felt material.

20. The cryogenic storage vessel of claim 18.

21. The felt material is polypropylene.

21. The cryogenic storage vessel of claim 20.

22. the first shell portion is a molded shell, the second shell portion includes a planar surface, and the first and second shell portions are rectangular. A cryogenic storage vessel according to any one of claims 1 to 21.

23. the rim has an extension, the extension forming part of a conduit between the interior volume and an exterior of the cryogenic storage vessel. A cryogenic storage vessel according to any one of claims 1-22.

24. an outer peripheral edge of the second flange of the second shell portion extends beyond an outer peripheral edge of the first flange of the first shell portion; 24. The cryogenic storage vessel of claim 23.

25. further comprising a through hole provided in at least one of the first flange and the second flange. A cryogenic storage vessel according to any one of claims 1-24.

26. The semiconductor device further includes at least one of an electronic sensor and a conductive circuit interposed between a first laminated layer of the first shell portion and a second laminated layer of the second shell portion. A cryogenic storage vessel according to any one of claims 1-25.

27. the electronic sensor is a temperature sensor; 27. The cryogenic storage vessel of claim 26.

28. The temperature sensor is selected from the group consisting of a thermistor, an RTD sensor, and a thermocouple.

28. The cryogenic storage vessel of claim 27.

29. a contact portion of the conductive circuit is exposed and / or accessible through the first and second laminated layers; A cryogenic storage vessel according to any one of claims 26-28.

30. further comprising a ferrite material disposed within the interior volume.

30. A cryogenic storage vessel according to any one of claims 1-29.

31. the ferrite material is a first sheet, the first sheet being laminated between a second sheet and a third sheet; 31. The cryogenic storage vessel of claim 30.

32. the first sheet and the second sheet are made of a polymer; 32. The cryogenic storage vessel of claim 31.

33. the interior volume includes a plurality of chambers; A cryogenic storage vessel according to any one of claims 1-32.

34. At least two of the plurality of chambers are independent of each other.

34. The cryogenic storage vessel of claim 33.

35. At least one of the plurality of chambers contains a liquid selected from the group consisting of a solution, a composition, a formulation, a particulate suspension, a virus suspension, a cell suspension, and a multicellular organism suspension.

34. The cryogenic storage vessel of claim 33.

36. At least two of the chambers are connected to each other.

34. The cryogenic storage vessel of claim 33.

37. the at least two chambers are connected to each other by a passageway selected from the group consisting of a conduit, an opening, a lumen, a conduit, a channel, and a tunnel; 37. The cryogenic storage vessel of claim 36.

38. 10. A method for preserving liquid in a cryogenic storage container for storing a frozen cell suspension according to claim 9, comprising the steps of: placing a recessed corner of the cryogenic storage vessel in a predetermined position; elevating the corner to a position higher than any other corner of the cryogenic storage vessel; penetrating the recess; inserting a filling needle into the pierced cavity; introducing liquid into the interior volume through a first portion of the fill needle while simultaneously removing displaced gas from the interior volume through a second portion of the fill needle; withdrawing the filling needle from the interior volume and the pierced cavity; closing the through cavity; and Including, method.

39. applying a protective cover over the blocked through recess.

39. The method of claim 38.

40. and further comprising removing a protective cover from the recess prior to penetrating the recess.

39. The method of claim 38.

41. and closing the pierced cavity includes inserting a plug into the pierced cavity.

39. The method of claim 38.

42. and further comprising the step of heat sealing the plug to the recess, wherein the interior volume is isolated from an exterior of the cryogenic storage vessel.

42. The method of claim 41.

43. 10. A method for withdrawing liquid from a cryogenic storage container for storing a frozen cell suspension according to claim 9, comprising the steps of: placing a first corner of the cryogenic storage vessel having a first recess in a predetermined position; elevating the corner to a position higher than the remaining corners of the cryogenic storage vessel; penetrating the recess with a vent tool; placing a second recess in a predetermined position at a second corner of the cryogenic storage vessel opposite the first corner of the cryogenic storage vessel; penetrating the second recess with an extraction tool; withdrawing the liquid from the cryogenic storage vessel; Including, method.

44. and further comprising the step of removing a protective cover from at least one of the first and second recesses prior to the penetrating step.

44. The method of claim 43.

45. the vent tool and the extraction tool are sterile; 44. The method of claim 43.

46. A method for thawing the coagulated contents of a cryogenic storage container for storing a frozen cell suspension according to any one of claims 30-32, comprising: the ferrite material is in contact with an inner surface of the first shell portion corresponding to the first major surface; The method comprises: clamping the cryogenic storage vessel between a first heater block and a second heater block, where the first heater block is in contact with the first major surface and the second heater block is in contact with the second major surface, the second heater block further comprising a magnetic source; increasing the temperature of the first and second heater blocks; activating the magnetic source; magnetically attracting the ferrite material to an inner surface of the second shell portion corresponding to the second major surface; displacing the solidified contents against an inner surface of the second shell portion through the ferrite material and the magnetic source; Including, method.

47. and decreasing the temperature of the first heater block when a liquid phase of the solidified contents appears within the cryogenic storage vessel.

47. The method of claim 46.

48. The magnetic source is selected from the group consisting of a permanent magnet and an electromagnet.

47. The method of claim 46.

49. further comprising removing the first and second heater blocks from the cryogenic storage vessel.

47. The method of claim 46.

50. A method for transferring cells from a cryopreservation medium to a restoration medium using a cryogenic storage container for storing a frozen cell suspension according to any one of claims 33 to 37, comprising: filling a first chamber of the cryogenic storage vessel with a cell suspension; filling a second chamber of the cryogenic storage vessel with a medium selected from the group consisting of a restoration medium and an injection medium; interconnecting the first and second chambers with a first passageway having a gate having a closed state and an open state, the closed state being changed to the open state by applying a centrifugal force to the cryogenic storage vessel; interconnecting the first and second chambers with a second passageway including a valve configured to permit passage of gas in response to the centrifugal force; applying a centrifugal force to the cryogenic storage vessel, wherein the cell suspension from the first chamber passes through a gate to the second chamber and gas in the second chamber passes through the valve to the first chamber; Including, method.

51. the second chamber is spaced apart from the first chamber along a centrifugal field vector of the cryogenic storage vessel.

51. The method of claim 50.

52. a portion of the second chamber is disposed away from the gate along the centrifugal field of the cryogenic storage vessel; 51. The method of claim 50.

53. positioning the gas in the second chamber at an end of the second chamber adjacent to the first chamber, the end of the second chamber running parallel to the first chamber and connecting the second chamber to the first chamber via the valve, such that only medium in the second chamber contacts the gate.

51. The method of claim 50.

54. The centrifugal force causes the cell suspension to pass through the gate into the second chamber and simultaneously causes air to pass through the valve into the first chamber.

51. The method of claim 50.

55. and further comprising the step of returning the gate to the closed state and closing the valve by reducing the centrifugal force.

51. The method of claim 50.

56. and further comprising the step of gently agitating the cell suspension and the medium in the second chamber.

56. The method of claim 55.

57. and removing the cell suspension and the medium from the second chamber through an outlet port of the cryogenic storage vessel.

57. The method of claim 56.

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