Cryopreservation device

Abstract

The present disclosure provides a vitrification straw for cryopreservation of biological samples. The vitrification straw comprises a holding means having a proximal end and a distal end, a tapered member positioned within the holding means, and a tip member located at the distal end. The tip member includes a curved support and a wavy tip. The wavy tip has a first end and a second end, forming a wavy cavity between them. The curved support and wavy tip are designed to ensure uniform cooling, prevent ice crystal formation, and enhance handling efficiency during the cryopreservation process.

Description

[0001] CRYOPRESERVATION DEVICE

[0002] TECHNICAL FIELD

[0003] [1] The present disclosure relates to cryopreservation devices, and more particularly to a vitrification straw, system, and method for enhanced cryopreservation of biological samples using a wavy tip and curved support design.

[0004] BACKGROUND

[0005] [2] Cryopreservation is a widely used technique for preserving biological samples, including cells, tissues, and embryos, by cooling them to very low temperatures. This process allows for long-term storage of valuable biological materials for research, medical applications, and conservation efforts. This process involves cooling biological samples to sub-zero temperatures, typically around -196°C using liquid nitrogen, to halt cellular metabolic processes and preserve the sample's viability for extended periods. The field of cryopreservation has seen significant advancements in recent years, with a focus on improving the survival rates of preserved samples and enhancing the efficiency of the freezing and thawing processes.

[0006] [3] Vitrification, a rapid cooling technique that transforms a liquid into a glass-like amorphous solid without the formation of ice crystals, has emerged as a preferred method for cryopreservation in many applications. This approach offers advantages over traditional slow freezing methods, particularly for delicate biological samples such as embryos and oocytes. Vitrification straws are specialized tools designed to facilitate this process, providing a means to hold and protect samples during the rapid cooling and subsequent storage in liquid nitrogen.

[0007] [4] The design of vitrification straws plays a crucial role in the success of the cryopreservation process. Traditional straws often consist of a simple tubular structure with a uniform cross-section. While functional, these designs can present challenges in achieving uniform cooling rates across the entire sample, potentially leading to inconsistent preservation outcomes.

[0008] [5] One of the primary technical problems associated with conventional vitrification straws is the difficulty in achieving rapid and uniform cooling. Uneven cooling rates can result in the formation of ice crystals in certain areas of the sample, compromising cellular integrity and reducing overall viability upon thawing. This issue is particularly critical for samples with complex structures or varying thermal properties.

[0009] [6] Another challenge faced in the field is the mechanical stress imposed on biological samples during the vitrification process. The rapid temperature changes and the physical handling of straws can subject delicate cells or tissues to potentially damaging forces. Existingstraw designs may not adequately protect samples from these stresses, leading to reduced survival rates or compromised sample quality.

[0010] [7] The loading and handling of samples into vitrification straws can also present difficulties for practitioners. Conventional straw designs may require precise manipulation techniques, increasing the risk of operator error and potentially compromising sample integrity. This can be particularly problematic in time-sensitive procedures or when dealing with limited or irreplaceable biological materials.

[0011] [8] Furthermore, the storage and retrieval of vitrified samples in liquid nitrogen containers pose additional challenges. Traditional straw designs may be susceptible to damage or deformation under the extreme cold conditions, potentially compromising the protective environment surrounding the preserved samples. This can lead to issues with long-term storage stability and sample recovery.

[0012] [9] The materials used in the construction of vitrification straws also present limitations. While various synthetic resins are employed, each material has its own set of drawbacks in terms of temperature resistance, mechanical strength, and biocompatibility. Finding a material that maintains its structural integrity and protective properties across the extreme temperature range involved in cryopreservation remains an ongoing challenge in the field.

[0013]

[0010] It has been appreciated that a vitrification straw with enhanced design features is needed that overcomes one or more of these problems.

[0014] SUMMARY

[0015]

[0011] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description.

[0016]

[0012] The present disclosure relates to an vitrification straw for cryopreservation of biological samples is provided. The vitrification straw includes a holding means having a proximal end and a distal end, an tapered member positioned within the holding means, and a tip member located at the distal end of the holding means, wherein the tip member comprises a curved support providing structural stability and a wavy tip extending from the curved support, the wavy tip having a wavy first end and a wavy second end, wherein the wavy tip forms a wavy cavity between the wavy first end and the wavy second end, and wherein the wavy tip is configured to promote uniform cooling of the biological samples during cryopreservation.

[0017]

[0013] This innovative design of the vitrification straw enhances the efficiency and effectiveness of cryopreservation processes for biological samples. The wavy tip promotes uniform cooling, which is critical for preventing ice crystal formation and maintaining cellviability. The curved support provides structural stability, ensuring the integrity of the straw during handling and storage under extreme cryogenic conditions.

[0018] BRIEF DESCRIPTION OF FIGURES

[0019]

[0014] The summary above, as well as the following detailed description of illustrative embodiments, is better understood when read in conjunction with the appended drawings. For the purpose of illustrating the present disclosure, exemplary constructions of the disclosure are shown in the drawings. However, the present disclosure is not limited to specific methods and instrumentalities disclosed herein. Moreover, those in the art will understand that the drawings are not to scale. Wherever possible, like elements have been indicated by identical numbers.

[0020]

[0015] FIG. 1 illustrates a perspective view of a vitrification straw, according to an embodiment of the present disclosure.

[0021]

[0016] FIG. 2 illustrates a perspective view of a tip member of the vitrification straw, according to an embodiment.

[0022]

[0017] FIG. 3 illustrates another perspective view of the tip member of the vitrification straw, according to an embodiment of the present disclosure.

[0023]

[0018] FIG. 4 illustrates a perspective view of the vitrification straw with a wavy cavity, according to an embodiment.

[0024]

[0019] FIG. 5 illustrates a top view of the tip member of the vitrification straw, according to an embodiment of the present disclosure.

[0025]

[0020] FIG. 6 illustrates a perspective view of a die means for manufacturing the vitrification straw, according to an embodiment.

[0026]

[0021] FIG. 7 illustrates side views of a vitrification straw (100) in uncapped and capped configurations.

[0027]

[0022] Common reference numerals are used throughout the figures to indicate similar features.

[0028] DETAILED DESCRIPTION

[0029]

[0023] The following detailed description illustrates embodiments of the present disclosure and ways in which they can be implemented. Although some modes of carrying out the present disclosure have been disclosed, those skilled in the art would recognize that other embodiments for carrying out or practicing the present disclosure are also possible

[0024] The present disclosure relates to cryopreservation devices and methods for preserving biological samples. In particular, embodiment of the disclosure may provide a vitrification straw with enhanced design features that may improve the efficiency and effectiveness of the cry opreservation process.

[0030]

[0025] In some embodiments, a vitrification straw may include a holding means, a tapered member, and a tip member. The tip member may comprise a curved support and a wavy tip. This configuration may provide several potential advantages for cryopreservation applications.

[0031]

[0026] The wavy tip design may promote more uniform cooling of biological samples during the vitrification process. In an embodiment, the undulating shape of the wavy tip may increase surface area and create turbulence in the cryoprotectant solution, which may modify the thermal behavior of the system. Cryoprotectants may lower the freezing point of water and alter the viscosity and thermal conductivity of the solution. These changes may influence the cooling and warming rates indirectly. However, the main role of cryoprotectants is to protect cells from damage due to ice formation during freezing, rather than to affect heat transfer efficiency directly. The curved support may provide structural stability to the tip member while also facilitating handling of the vitrification straw.

[0032]

[0027] Additionally, the wavy cavity formed within the tip member may allow for improved sample loading and retrieval. The cavity's shape may help retain samples securely during the rapid cooling process while still allowing for efficient heat exchange.

[0033]

[0028] Various manufacturing methods for producing the vitrification straw are also described herein. In an embodiment, extrusion techniques utilizing specialized die means may be employed to form the wavy tip and curved support features of the straw.

[0034]

[0029] The following detailed description provides further information on the structure, function, manufacture, and use of the vitrification straw according to various embodiments. While specific examples are discussed, it should be understood that these are provided for illustrative purposes and that various modifications are possible within the scope of the disclosure.

[0035]

[0030] The present disclosure describes a vitrification straw 100, as illustrated in FIG.l, which is designed for the cry opreservation of biological samples. The vitrification straw 100 comprises a holding means 101, a tapered member 102, and a tip member 103, each contributing to the functionality and efficiency of the cryopreservation process.

[0036]

[0031] The holding means 101 forms the main body of the vitrification straw 100 and extends from a proximal end 100a to a distal end 100b. The vitrification straw 100 may beconstructed from a variety of materials suitable for cryopreservation applications. In an embodiment, the vitrification straw 100 may be made of polymeric materials including, but not limited to polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyurethane (PU) or polycarbonate (PC). These materials may offer advantages in terms of flexibility, chemical resistance, and thermal stability at low temperatures. In a specific embodiment, the vitrification straw 100 is constructed from polycarbonate, a material chosen for its durability and resistance to low temperatures, which are critical in cryopreservation environments.

[0037]

[0032] The proximal end 100a of the vitrification straw 100 is designed to facilitate easy handling and insertion during the cryopreservation process, while the distal end 100b connects to the tip member 103, which is critical for the specific functionalities of cry opreservation.

[0038]

[0033] Within the holding means 101, a tapered member 102 is positioned to extend along the length of the holding means 101. This tapered member 102 provides structural support to the vitrification straw 100, ensuring that it maintains its integrity during the vitrification process.

[0039]

[0034] The tip member 103, located at the distal end 100b of the vitrification straw 100 of the holding means 101. The tip member 103 includes features that are crucial for the effective handling and storage of biological samples. These features are designed to optimize the cooling rates and ensure uniform temperature distribution across the biological samples, thereby minimizing the risk of ice crystal formation which can damage the samples.

[0040]

[0035] Overall, the structure of the vitrification straw 100, including the holding means 101, tapered member 102, and tip member 103, is engineered to provide an optimal environment for the cry opreservation of sensitive biological materials. The use of polycarbonate in the construction of the holding means 101 ensures that the vitrification straw 100 is capable of withstanding the harsh conditions of cryopreservation while maintaining the viability and integrity of the biological samples contained within.

[0041]

[0036] The tip member 103, as illustrated in FIG.2, FIG.3, and FIG.5, includes a curved support 103a and a wavy tip 103b, each contributing uniquely to the overall performance of the vitrification straw 100.

[0042]

[0037] A curved support 103 a is incorporated at the base of the tip member 103. This curved support 103a provides a stable and secure environment for the placement of embryos during the cryopreservation process. The design of the curved support 103a ensures that embryos are held firmly in place, minimizing the risk of displacement or damage during storage and handling.

[0038] In an embodiment, the curved support 103a may be integrally connected with the wavy tip 103b and the tip member 103 to form a cohesive structure. The curved support 103a may transition smoothly into the wavy tip 103b, creating a continuous surface.

[0043]

[0039] Adjacent to the curved support 103a is a wavy tip 103b, which is characterized by its undulating shape. The wavy tip 103b extends outward from the curved support 103a, as depicted in FIG.3 and FIG.5.

[0044]

[0040] In an embodiment, the wavy tip 103b includes a first end 103bl and a second end 103b2 that may connect to the curved support 103a. This configuration may create a continuous structure that from the curved support 103a to the undulating surface of the wavy tip 103b. As shown in FIG.3 and FIG.5, the wavy tip 103b may include multiple peaks and troughs along its length. In an embodiment, the wavy tip 103b may have a sinusoidal shape, a zigzag pattern, or other undulating geometries. The amplitude and frequency of the waves in the wavy tip 103b may be varied to achieve different effects. Additionally, the wavy tip 103b may incorporate asymmetric wave patterns or graduated wave sizes along its length in an embodiments.

[0045]

[0041] In an embodiment, the design of the tip member 103 allows for the loading of embryos from either side of the tip, providing flexibility and ease of use in various laboratory settings. This adaptability is particularly beneficial in environments where precision and efficiency are paramount.

[0046]

[0042] In an embodiment, the curved support 103a may have a different shape. The shape of the curved support 103a may include, but is not limited to, a C shape, a U shape, or a V shape. In an embodiment, the curved support 103a may incorporate a combination of shapes or asymmetric designs.

[0047]

[0043] The curved support 103a may also have other curved or angled configurations. In an embodiment, the curved support 103a may incorporate a combination of shapes or asymmetric designs.

[0048]

[0044] The vitrification straw 100 includes a wavy cavity 103b3 within the wavy tip 103b, as illustrated in FIG.4. The wavy cavity 103b3 extends along the length of the tip member 103, forming a channel -like structure that follows the undulating contours of the wavy tip 103b.

[0049]

[0045] The wavy cavity 103b3 is configured to hold biological samples during the cryopreservation process. The curved nature of the cavity may help prevent displacement of cells during handling and storage. As the wavy cavity 103b3 follows the contours of the wavy tip 103b, cells may be more securely held in place compared to a straight cavity.

[0046] The wavy tip 103b, featured prominently in the design, introduces a novel approach to managing the cooling process. The undulating shape of the wavy tip 103b increases the surface area in contact with the cryoprotectant solution.

[0050]

[0047] Moreover, the wavy cavity 103b3, formed within the wavy tip 103b, plays an essential role in the cryopreservation process. This cavity allows for the secure placement of biological samples, ensuring that they are evenly exposed to the cooling effects of the cryoprotectant solution. The design of the wavy cavity 103b3 follows the contours of the wavy tip 103b, which helps in maintaining a consistent temperature gradient throughout the sample, further aiding in the prevention of ice crystal formation.

[0051]

[0048] The interaction between the curved support 103a and the wavy tip 103b is particularly important. The curved support 103a not only stabilizes the wavy tip 103b but also ensures smooth transition between different parts of the tip member 103, maintaining a continuous flow of the cryoprotectant solution without any disruptions. This continuity is vital for achieving the desired uniformity in cooling, which is critical for the success of the cryopreservation process.

[0052]

[0049] The tip member 103 of the vitrification straw 100 may have specific dimensions that contribute to its functionality in the cry opreservation process. The length of the tip member 103 may range from about 05 mm to 40 mm, providing sufficient space for sample loading and manipulation. In an specific embodiment, length of the tip member 103 is 20 mm. The width of the tip member 103 may vary along its length due to the wavy design, with an average width potentially ranging from about 1.5 mm to 4.0 mm. In an specific embodiment width of the tip member 103 is 2.5 mm.

[0053]

[0050] The thickness of the tip member 103 walls may be in the range of 0.10 mm to 0.50 mm, balancing structural integrity with thermal conductivity. In an specific embodiment thickness of the tip member 103 walls is 0.15 mm. The wavy cavity 103b3 within the tip may have a depth of 0.3 mm to 0.6 mm, allowing for adequate sample volume while maintaining proximity to the cooling surface.

[0054]

[0051] In an embodiment, the curved support 103a may extend 1 mm to 2.5 mm in height from the main body of the tip, The amplitude of the waves in the wavy tip 103b may range from 1.2 mm to 2 mm in width. These dimensions may be adjusted based on specific application requirements, manufacturing constraints, and optimization for different types of biological samples.

[0052] The manufacturing process of the vitrification straw 100 involves specialized equipment and techniques to create the unique features of the straw, particularly the wavy tip 103b and curved support 103a. A key component in this process is a die means 200, which includes a wavy shape channel 201, as illustrated in FIG.6.

[0055]

[0053] The process begins with the preparation of materials. Polymer pellets, such as polypropylene, polyurethane, Polyethylene Terephthalate (PET), Polycarbonate (PC), Polymethyl Methacrylate (PMMA), Polyvinyl Chloride (PVC), are selected based on their thermal properties and biocompatibility. Additives like stabilizers, UV protectants, and plasticizers are blended with the polymer pellets to enhance the properties of the final product. In a specific embodiment, the vitrification straw 100 is constructed from Polyethylene Terephthalate (PET), which may provide suitable properties for cryopreservation applications. The die means 200 is engineered with specific channels that create the desired shapes of the wavy tip 103b and the curved support 103a. As shown in FIG.6, the die means 200 includes a wavy shape channel 201, which is designed to form the undulating structure of the wavy tip 103b. The wavy shape channel 201 is positioned on the angled face of the die means 200, allowing for the controlled extrusion of the material.

[0056]

[0054] During the manufacturing process, the blended polymer material is passed through the die means 200. As the material flows through the wavy shape channel 201, the unique geometry of the channel shapes the material into the desired wavy tip 103b and curved support 103a configurations. The design of the wavy shape channel 201 ensures that the material fills all areas properly, creating the distinctive features of the vitrification straw 100.

[0057]

[0055] Precise temperature control is maintained in the die means 200 to prevent premature solidification of the polymer material. This temperature control is crucial for ensuring optimal material flow and shaping. The carefully regulated temperature allows the material to maintain its fluidity as the wavy shape channel 201 molds the wavy tip 103b and curved support 103a.

[0058]

[0056] As the molten polymer exits the die means 200, the extruded material takes on the form of the vitrification straw 100, complete with the wavy tip 103b and curved support 103 a. The extruded straw then undergoes cooling and any necessary post-processing steps to finalize the product.

[0059]

[0057] This manufacturing process, utilizing the die means 200 and wavy shape channel 201, allows for the consistent production of vitrification straws 100 with the specialized features required for effective cry opreservation of biological samples.

[0058] FIG. 7 illustrates side views of a vitrification straw 100 in two configurations: uncapped and capped. The lower portion of FIG. 7 depicts the vitrification straw 100 with a protective cap 300 in place. The protective cap 300 is fitted over the tip member 103, enclosing and safeguarding it. This cap may provide protection during handling, storage, and transportation of the vitrification straw (100). In some embodiments, the vitrification straw 100 may include a protective cover that extends beyond a simple cap design. This protective cover may take various forms such as a sleeve, sheath, or enclosure that partially or fully surrounds the vitrification straw 100. The cover may be made from materials such as polyethylene (PE), polycarbonate (PC), polymethyl methacrylate (PMMA), acetal (POM). The cover may be designed to be removable, or it may be a single-use disposable component.

[0060]

[0059] The vitrification process for cryopreservation of biological samples using the vitrification straw 100 may involve several steps to ensure optimal preservation and viability of the samples.

[0061]

[0060] Initially, the biological sample may be prepared in a suitable culture medium. The sample may then be gradually exposed to a series of cryoprotectant solutions with increasing concentrations. This stepwise approach may help minimize osmotic shock to the cells.

[0062]

[0061] The sample may then be carefully loaded into the wavy cavity 103b3 of the tip member 103. The undulating shape of the wavy tip 103b may help distribute the sample evenly and securely hold it in place. The curved support 103a may provide additional stability during handling.

[0063]

[0062] Rapid cooling may then be achieved by plunging the loaded vitrification straw 100 directly into liquid nitrogen. This may allow the sample to transition quickly to a vitrified state without ice crystal formation. The cooling may be extremely rapid, typically within a few seconds to prevent ice crystal formation, which can damage the cell structure. This rapid cooling may form a vitrified state, where the water in the sample solidifies into a glass-like structure without crystallizing

[0064]

[0063] For long-term storage, the vitrification straw 100 containing the vitrified sample may be kept submerged in liquid nitrogen at approximately -196°C.

[0065]

[0064] When the sample is needed, the vitrification straw 100 may be rapidly warmed, potentially in a warm water bath. Following warming, the sample may be carefully removed from the tip member 103 and rehydrated by gradually diluting the cryoprotectants.

[0066]

[0065] Throughout this process, the specialized features of the vitrification straw 100, such as the wavy tip 103b and curved support 103a, may contribute to maintaining sample integrity and optimizing cooling / warming rates. The overall design may allow for precise control oftemperatures and cryoprotectant exposure, which may be important factors in successful vitrification outcomes.

[0067]

[0066] Features of any of the examples or embodiments outlined above may be combined to create additional examples or embodiments without losing the intended effect. It should be understood that the description of an embodiment or example provided above is by way of example only, and various modifications could be made by one skilled in the art. Furthermore, one skilled in the art will recognize that numerous further modifications and combinations of various embodiments are possible. Accordingly, the described embodiments are intended to encompass all such alterations, modifications, and variations that fall within the scope of the appended claims.

Claims

We Claim,1. A vitrification straw (100) for cry opreservation of biological samples, comprising:a holding means (101) having a proximal end (100a) and a distal end (100b);an tapered member (102) positioned within the holding means (101); anda tip member (103) located at the distal end (100b) of the holding means (101), wherein the tip member (103) comprises:a curved support (103a) providing structural stability; anda wavy tip (103b) extending from the curved support (103a), the wavy tip (103b) having a first end (103b 1) and a second end (103b2), wherein the wavy tip (103b) is configured to promote uniform cooling of the biological samples during cryopreservation.

2. The vitrification straw (100) as claimed in claim 1, wherein the vitrification straw (100) made of polymeric materials such as polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyurethane (PU).

3. The vitrification straw (100) as claimed in claim 1, wherein the vitrification straw (100) is constructed from polycarbonate, which may provide durability and resistance to low temperatures encountered during cryopreservation.

4. The vitrification straw (100) as claimed in claim 1, wherein the wavy tip (103b) includes multiple peaks and troughs along its length.

5. The vitrification straw (100) as claimed in claim 1, wherein the wavy tip (103b) comprises a shape selected from the group consisting of a sinusoidal shape, a zigzag pattern, and other undulating geometries.

6. The vitrification straw (100) as claimed in claim 1, wherein the wavy tip (103b) forms a wavy cavity (103b3) between the first end (103b 1) and the second end (103b2).

7. The vitrification straw (100) as claimed in claim 6, wherein the wavy cavity (103b3) extends along the length of the tip member (103).

8. The vitrification straw (100) as claimed in claim 6 or 7, wherein the wavy cavity (103b3) forms a channel-like structure that follows the undulating contours of the wavy tip (103b).

9. The vitrification straw (100) as claimed in claim 1, wherein the curved support (103a) comprises a shape selected from the group consisting of a C shape, a U shape, and a V shape.

10. A method of manufacturing a vitrification straw (100) for cryopreservation of biological samples, the method comprising:selecting a polymer material from a group consisting of polypropylene, polyurethane, polyethylene terephthalate, polycarbonate, polymethyl methacrylate, polyvinyl chloride; blending the selected polymer material with additives to form a polymer blend, wherein the additives may include stabilizers, UV protectants, and plasticizers;providing a die means (200) comprising a wavy shape channel (201);passing the polymer blend through the die means (200), wherein the wavy shape channel (201) shapes the polymer blend to form a tip member (103) comprising a curved support (103a) and a wavy tip (103b); and cooling the shaped polymer blend to produce the vitrification straw (100).

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