Device for cultivating and transporting biological tissue

Abstract

A two-part container for cultivating and / or transporting biological tissue and associated methods of use are disclosed. The container may include a lid having a first sidewall extending around a perimeter of the lid, and the first sidewall may include a first thread pattern. The container may include a base having a second sidewall extending around a perimeter of the base, and the second sidewall may include a second thread pattern corresponding in size and shape to the first thread pattern of the lid. The container may include an O-ring disposed in an annular channel formed in the second sidewall such that, in a tightened position, the first sidewall of the lid compresses the O-ring against the second sidewall of the base thereby forming a protected storage space. Disclosed containers may include flanges that define a receiving cavity suitable for stacking a plurality of two-part containers.

Description

DEVICE FOR CULTIVATING AND TRANSPORTING BIOLOGICAL TISSUERELATED APPLICATION S)

[0001] This application claims the benefit of U.S. Patent Application No. 63 / 541,304, filed on September 29, 2023. The entire teachings of the above application are incorporated herein by reference.FIELD

[0002] The present technology is generally related to a two-part container device for cultivating and / or transporting biological tissue.BACKGROUND

[0003] Biological tissue is often cultivated in a standard medical grade petri dish and then transported to a medical facility for use in a surgical procedure. In many instances, biological tissue is cultivated on surgical sponges in standard medical grade petri dishes. Once the biological tissue is cultivated it is often hard to transfer from a first petri dish used for cultivation to a secondary petri dish or other container used for transportation.

[0004] Typically, the biological tissue is transported in a petri dish which has a base section and a loose-fitting lid. The inherent properties of a standard petri dish subject the biological tissue to various contamination risks during the delivery and transportation of the biological tissue, e.g., to secondary petri dishes and / or from a culture site to a medical facility. Although this may be sufficient means for transportation of some biological tissues, it is not acceptable for at risk biological tissues to be implanted into patients, such as thymus tissue. For example, a loose-fitting lid may allow microbial intrusion, unwanted atmosphere, and other types of contaminants to enter the petri dish and contaminate the biological tissue. Therefore, a need exists for a container that is sufficient for use as a means to culture biological tissue as well as a means to transport the cultured biological tissue in the same container.SUMMARY

[0005] The techniques of this disclosure generally relate to a medical grade container formed of a base portion and a lid portion, referred to herein also as a two-part container.The lid portion may threadedly secure to the base portion and provide a sealed interior by virtue of the threads and the compression of a medical grade O-ring.

[0006] In designing the medical grade containers disclosed herein, a first consideration was ensuring the design would prevent ingress of contaminants from the environment to the biological ti ssue stored inside of the container during transport (e.g., during transport of the biological tissue from a place of origin to a surgical center such as a hospital). An example biological tissue may be a thymus specimen although other types of biological tissue are contemplated. In various embodiments, a thymus specimen may be sourced from a donor or donors for subsequent culturing and implantation in a recipient. In at least one example embodiment, an individual may be the source of a tissue donation that is cultured and later implanted in that same individual. For example, the individual is both the source of tissue donation and the recipient of a tissue sample.

[0007] In various embodiments, an O-ring may be provided to prevent the egress of moisture which is used to ensure a hospitable environment for the biological tissue.Thus, the design disclosed herein includes threading a lid portion onto the base portion and using a medical grade O-ring that is capable of maintaining the viability of a biological tissue for extended transportation periods, including, but not limited to, up to about 72 hours to about 96 hours. A second consideration i n designing the medical grade containers disclosed herein was ensuring the quality of the biological tissue would be maintained, by evaluating the appropriate environment for the biological tissue during the delivery process and ensuring the container could provide that environment to ensure that the biological tissue stored inside of the container remained viable throughout the transportation phase. Another factor in ensuring quality of the biological tissue was to ensure use of appropriate materials of construction for use in a medical context. For example, the materials used in the containers disclosed herein are preferably durable and capable of withstanding a range of different sterilization methods over repeated use cycles. Such example sterilization methods may include the use of ethylene oxide and e-beam sterilization methods, which are known to degrade conventional containers. A third consideration in designing the medical grade containers disclosed herein was ensuring the biological tissue could be easily removed from the container in the surgical suite during the surgical implantation procedure such that the biological tissue would not be damaged during the removal phase. In this sense, the design of the containers disclosed herein includes a relatively wide opening to the interior storage cavity’ and relatively short walls to provide easy access to the biological tissue with standard tools used in surgery.Additionally, the interior of the container was designed with several internal partitions to separate a plurality of sponges supporting the biological tissue to keep the sponges from shifting around during transportation and silo the sponges off from one another. Another aspect is to provide a secondary storage package for stacking a plurality of the individual two-part containers disclosed herein.

[0008] In one aspect, the present disclosure provides a two-part container for cultivating and / or transporting biological tissue. As used herein, the terms cultivating and culturing are used interchangeably. The container may include a lid having a first sidewall extending around a perimeter of the lid, and the first sidewall may include a first thread pattern. The container may include a base having a second sidewall extending around a perimeter of the base, and the second sidewall may include a second thread pattern corresponding in size and shape to the first thread pattern of the lid. The container may include an O-ring disposed in an annular channel formed in the second sidewall such that, in a tightened position, the first sidewall of the lid compresses the O-ring against the second sidewall of the base thereby forming a protected storage space.

[0009] Another aspect of the disclosure includes a method of using and sterilizing a two- part container for cultivating and / or transporting biological tissue. The method may include the step of providing a two-part container, including a lid having a first sidewall extending around a perimeter of the lid, the first sidewall including a first thread pattern, and a base having a second sidewall extending around a perimeter of the base, the second sidewall including a second thread pattern corresponding in size and shape to the first thread pattern of the lid. The base may include a floor having an interior surface for supporting biological tissue having a plurality of walls configured to partition the interior surface into a plurality of receiving areas, and the plurality of walls are discontinuous such that moisture may be dispersed between the plurality of receiving areas. The two-part container may further include an O-ring disposed in an annular channel formed in the second sidewall. The method may further include the step of sterilizing the two-part container via an irradiation technique and culturing a plurality of biological tissue samples in the base for at least seven days. Each biological tissue sample may be disposed on a medical grade sponge in a respective receiving area. The method may further include the step of securing, after the culturing step, the lid to the base by engaging the first thread pattern and the second thread pattern thereby compressing the O-ring between the first sidewall and second sidewall to form a protected storage space, and the storage space may contain a sufficient volume of oxygen to support the viability of each biological tissue sample for at least 72 hours. The method may furtherinclude the step of transporting the two-part container to a different location from where the culturing step took place.

[0010] According to another aspect, a method of using, sterilizing, and re-using a plurality of two-part containers for cultivating and / or transporting biological tissue is disclosed. Each two-part container may be stacked on top of an adjacent two-part container and transported in the stacked configuration.

[0011] The details of one or more aspects of the disclosure are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the techniques described in this disclosure will be apparent from the description and drawings, and from the claims.BRIEF DESCRIPTION OF DRAWINGS

[0012] FIG. 1 is a perspective view of an example two-part container shown in skeleton view.

[0013] FIG. 2 is a side profile view of the two-part container shown in FIG. 1 in skeleton view.

[0014] FIG. 3 is a top-down view of the two-part container shown in FIG. 1 in skeleton view.

[0015] FIG. 4 is a perspective view of a base portion of the two-part container shown in FIG. 1 in skeleton view.

[0016] FIG. 5 is a perspective view of a lid portion of the two-part container shown in FIG. 1 in skeleton view.

[0017] FIG. 6 is a cross-section view of the two-part container taken through section 6-6 of FIG. 3.

[0018] FIG. 7 is a perspective view of an example manufactured two-part container with the lid portion unscrewed from the base portion for ease of understanding.

[0019] FIG. 8 is a side profile view of an example carrier and a plurality of example stacked two-part containers.

[0020] FIG. 9 is a graph showing the concentration of CCL21 protein in pg / ml at different time points during and after shipment for control and shipped thymus tissue cultures (PD = post day shipping).DETAILED DESCRIPTION

[0021] Embodiments of the present disclosure relate generally, for example, to medical grade containers for the culturing and transportation of biological materials, and more particularly, to two-part containers. Embodiments of the devices and methods are described below with reference to the Figures.

[0022] The following discussion omits or only briefly describes certain components, features and functionality related to medical implants, installation tools, and associated surgical techniques, which are apparent to those of ordinary skill in the art. It is noted that various embodiments are described in detail with reference to the drawings, in which like reference numerals represent like parts and assemblies throughout the several views, where possible. Reference to various embodiments does not limit the scope of the claims appended hereto because the embodiments are examples of the inventive concepts described herein. Additionally, any example(s) set forth in this specification are intended to be non-limiting and set forth some of the many possible embodiments applicable to the appended claims. Further, particular features described herein can be used in combination with other described features in each of the various possible combinations and permutations unless the context or other statements clearly indicate otherwise.

[0023] Terms such as “same,” “equal,” “planar,” “coplanar,” “parallel,” “perpendicular,” etc. as used herein are intended to encompass a meaning of exactly the same while also including variations that may occur, for example, due to manufacturing processes. The term “substantially” may be used herein to emphasize this meaning, particularly when the described embodiment has the same or nearly the same functionality or characteristic, unless the context or other statements clearly indicate otherwise. The term “about” may also be used herein to emphasize this meaning and if a value and / or a range of values is provided in the specification or claims with the modifier “about” a meaning of + / - ten percent (10%) to those provided values are encompassed by the meaning of “about,” unless the context clearly indicates otherwise.

[0024] Components of the disclosed embodiments can be fabricated from biologically acceptable materials suitable for medical applications, including metals, synthetic polymers, ceramics and bone material and / or their composites. For example, the components, individually or collectively, can be fabricated from materials comprising thermoplastics such as polyaryletherketone (PAEK) including polyetheretherketone (PEEK), polyetherketoneketone (PEKK) and polyetherketone (PEK), carbon-PEEK composites,PEEK-BaS04 polymeric rubbers, polyethylene terephthalate (PET), fabric, silicone, polyurethane, silicone-polyurethane copolymers, polymeric rubbers, polyolefin rubbers, semi-rigid and rigid materials, elastomers, rubbers, thermoplastic elastomers, thermoset elastomers, elastomeric composites, rigid polymers including polyphenylene, polyamide, polyimide, polyetherimide, polyethylene, epoxy, and their combinations.

[0025] FIG. l is a perspective view of an example two-part container 100 shown in skeleton view. The primary components of container 100 are base 10, lid 20, and O-ring 30. In practice, the base 10 and lid 20 are securely tightened together with the O-ring being positioned between an outside surface of the base 10 and inside lip (or seating ring) of the lid 20 to provide a sealed environment on the inside of the container 100. In the example embodiment, base 10 and lid 20 have a substantially circular shape and are corresponding in size and shape. In various embodiments, the base 10 and lid 20 may comprise substantially planar exposed exterior surfaces and substantially planar interior surfaces as shown in the corresponding drawings.

[0026] FIG. 2 is a side profile view of the two-part container 100 shown in FIG. 1 in skeleton view. As illustrated, base 10 may include a pair of oppositely disposed flanges 11 on a lower end. Each flange 11 may extend for a distance below the exposed central lower surface of the base portion thereby forming a nesting cavity 18 between the opposite flanges 11. The nesting cavity may have a size and shape sufficient to allow the stacking of containers 100 (see FIG. 8).

[0027] FIG. 3 is a top-down view of the two-part container 100 shown in skeleton view. In the example embodiment, the interior of the container 100 is illustrated. The interior of the container 100 may be substantially defined by the interior perimeter of the base 10. For example, the sidewalls of base 10 may delimit the perimeter of the base 10. The interior of the base 10 may include a plurality of walls 12, 13 and a plurality of receiving areas 14, 15. In the example embodiment, a first receiving area 14 is defined by a pair of long walls 12 and a pair of short walls 13. Similarly, a second receiving area 15 is defined by a pair of long walls 12 and a pair of short walls 13. In various embodiments, the walls 12, 13 may be spaced apart from one another to allow for the free exchange of atmosphere between receiving areas as well as liquids and humidity between receiving areas. In other embodiments (not illustrated), the walls may be closed to prevent the exchange of atmosphere and liquids. In use, a sponge for culturing and supporting biological tissue may be disposed in a respective receiving area 14, 15. The corresponding plurality of walls 12, 13 may have the added benefit of preventing the sponges and biological tissue from moving around duringtransportation. Additionally, the floor of the base 10 may include various label markings to allow for the labeling and individual tracking of various specimens (see FIG. 3 label markings divided in four quadrants A, B, C, and D). In various embodiments, the floor of the base 10 may remain uncoated because coatings have been shown to have a potential to contaminate at risk biological tissue. Hence, a moist sponge for cultivating the biological tissue has been determined an effective means to support the biological tissue and to facilitate its removal.

[0028] FIG. 4 is a perspective view of the base 10 of the two-part container 100 shown in skeleton view and FIG. 5 is a perspective view of the lid 20. In the example illustration, the first and second receiving areas 14, and 15 are shown separated from one another by the corresponding plurality of walls 12, 13. Additionally, the O-ring 30 is shown nested in a recessed ring like cavity having a size and shape corresponding to a size and shape of the Ciring 30. In this example, the O-ring 30 may be a durable medical grade silicon O-ring which is shown to have superior sealing ability as well as withstand E-beam sterilization methods and repeated use. In some embodiments, the O-ring may be a medical grade silicon O-ring and in other embodiments the O-ring may be a food grade O-ring. Additionally, in some embodiments the O-ring may be non-coated or coated with, for example, perylene. Since an uncoated O-ring 30 may be permeable to air and given the size of the O-ring 30 relative to the volume of the dish, the permeability of the O-ring may be an important design consideration in instances where some gas exchange is desired vs. instances where no gas exchange is desired.

[0029] An outside sidewall of the base 10 may include a first thread pattern 16. Thread pattern 16 may have a run out portion or a stop surface to facilitate preventing and / or suppressing the overtightening of the lid 20 with respect to the base 10. Additionally, the flanges 11 may have a plurality of ridges 19 spaced at even or irregular intervals around the circumference of the exposed sidewalls of the base 10. Similarly, the lid 20 may have a plurality of ridges 29 spaced at even or irregular intervals around the circumference of the exposed sidewalls of the lid 20. In use, the container 100 has a relatively wide mouth and short sidewalls 37, 38 and the ridges 19, 29 make the container 100 relatively easy to open and close. In some embodiments, a first ridge 19 of the base portion may function as a first indicator and a second ridge 29 of the lid portion may function as a second indicator that allows a user to know when the lid 20 is sufficiently tightened to the base 10 that a sufficient amount of pressure is applied to the O-ring 30 and a sufficient seal is formed. Other types ofindicator markings such as arrows, notches, etc. (not illustrated) are also contemplated for this purpose.

[0030] FIG. 6 is a cross section view of the two-part container taken through section 6-6 of FIG. 3. As shown, lid 20 includes a thread pattern 26 on an interior surface of the sidewalls 38 that is mated with a corresponding thread pattern 16 on the exterior surface of sidewalls 37. Additionally, the exterior of sidewall 37 of the base 10 includes a lip 32 and a recessed cavity 31 that circumscribes the exterior surface of the base 10 sidewalls 37. For example, the recessed cavity may be an annular ring like cavity. The recessed cavity 31 and lip 32 are sized to receive the O-ring 30. In use, when the lid 20 is rotatably secured to the base 10 the interior surface of the lid 20 sidewalls 38 compresses the O-ring 30 into the recessed cavity 31 thereby forming a seal that is sufficient to prevent the microbial ingress of contaminants without resulting in an excessive torque to open the container 100. Preliminary testing has shown that the torque required to close the container 100 ranged from about 21 to 45 in-lb. and the torque to open the container 100 ranged from about 20 to 37 in-lb. In various embodiments, the seal may also prevent the exchange of Oxygen (which is required to keep the biological tissue alive) although the available volume inside of the protected storage space 39 is sufficient to maintain the viability of the biological tissue for at least 72 hours and upwards of 144 hours. In at least one embodiment, the available volume inside of the protected storage space 39 is sufficient to maintain the viability of the biological tissue for about 72 hours to about 96 hours.

[0031] In this way, the two-part container 100 forms a protected storage space 39 that is substantially closed off to the outside environment. Protected storage space 39 may be suitable for preventing the ingress and egress of microbial structures, gases, and liquids thereby protecting sensitive biological tissue stored inside of protected storage space 39. Additionally, other features such as the plurality of walls 12, 13 are shown inside the interior of protected storage space 39 to help maintain sponges and biological tissue in place during transport. In some embodiments, container 100 may be configured to allow some gas exchange to allow for culturing of biological tissue and the preservation of the biological tissue during transportation (not illustrated). This may be accomplished in a variety of ways, e.g., choosing an O-ring 30 that allows for an appropriate amount of gas exchange while still substantially sealing off the protected storage space 39 from microbial ingress. Other embodiments may include a valve (not illustrated) in the lid 20 or base 10 that allows for gas exchange, purging, or pressure release.

[0032] It should be noted that in various embodiments during the culturing process the lid 20 may not be completely tightened such that the free exchange of atmosphere is allowed. In some of these embodiments, the culturing process may take place in an oxygen rich environment. In other methods of culturing, the lid 20 may be loosely installed to allow for the free exchange of atmosphere or an alternate lid from a conventional petri dish may be used with the base 10 during the culturing phase. However, during transportation the lid 20 should be sufficiently tightened to protect the cultured at risk biological tissue stored within the container 100.

[0033] In one embodiment, the biological tissue may be thymus tissue. In one embodiment, the thymus tissue is fresh thymus tissue. In one embodiment, the biological tissue is cultured thymus tissue that has undergone a cultivation process. In one embodiment, the thymus tissue has undergone short-term storage at room temperature prior to culturing. In one embodiment, the biological tissue is partially T cell depleted thymus tissue. In one embodiment, the thymus tissue has undergone cry opreservation prior to culturing. In embodiments, the biological tissue is a cultured thymus tissue has that been generated using any of the cultivation methods disclosed in PCT application No. PCT / US2019 / 019137 (PCT Publication No. WO2019165197), PCT application No. PCT / US2020 / 046341 (PCT Publication No. W02021034650), and / or PCT application No. PCT / IB2023 / 058415, all which are incorporated herein in their entireties.

[0034] In some embodiments, the biological tissue is a skin, liver, or lung tissue. In various embodiments, the biological tissue may be a tissue slice. In embodiments, the biological tissue is suitable for implantation into a subject. In one embodiment, the subject is a human. In another embodiment, the subject is a non-human primate. In some embodiments, the subject is a rodent (such as a mouse, a rat, rabbit, etc.) or a dog. In one embodiment, the subject is an infant.

[0035] The containers disclosed herein may be used to culture biological tissue. In embodiments, the biological tissue is cultivated in a container disclosed herein for a period of few hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, or about 21 days. In certain embodiments, the culturing is for a period of about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, or about 21 days. In certain embodiments, the culturing is for a period of5-6 days, 5-7 days, 5-8 days, 5-9 days, 5-10 days, 6 to 7 days, 6 to 8 days, 6 to 9 days, 6 to 10 days, 6 to 11 days, 6 to 12 days, 6 to 21 days, 7 to 21 days, 8 to 21 days, 9 to 21 days, 10 to 21 days, 11 to 21 days, 12 to 21 days, 13 to 21 days, 14 to 21 days, 15 to 21 days, 16 to 21 days, 17 to 21 days, 18 to 21 days, 19 to 21 days, or 20 to 21 days. In embodiments, the biological tissue is cultured in a container disclosed herein for a period of about 6 days to about 21 days. In embodiments, the biological tissue is cultured in a container disclosed herein for a period of about 7 days to about 19 days. In an embodiment in which the biological tissue is a thymus tissue, a preferred cultivation period is at least about 12 days. However, a cultivation period for thymus tissue may range from about 5 days up to about 11 days.

[0036] Media for culturing biological tissues are known to a person skilled in the art. In embodiments, the media comprises HAMS F12 Nutrient Mixture, N-2- hydroxyethylpiperazine-N’-2-ethanesulfonic acid (HEPES) buffer, L-Glutamine, and Fetal Bovine Serum (FBS).

[0037] The containers disclosed herein may be used to transport biological tissue. In embodiments, the containers provided herein support viability of the biological tissue and / or suitability of the biological tissue for implantation for at least about 12 hours, at least about 24 hours, at least about 48 hours, at least about 72 hours, at least about 96 hours, at least about 120 hours, or at least about 144 hours. Methods of testing the viability of biological tissue and / or suitability' of the biological tissue for implantation are known in the art. In embodiments, viabi lity of thymus ti ssues or thymus-derived tissues can be determined using a method, one or more biomarkers, and / or one or more histological markers disclosed in PCT application No. PCT / US2019 / 019137 (PCT Publication No. WO2019165197), PCT application No. PCT / US2020 / 046341 (PCT Publication No. W02021034650), and / or PCT application No. PCT / IB2023 / 058415.

[0038] FIG. 7 is a perspective view of a manufactured two-part container 100 with the lid 20 unscrewed from the base 10 for ease of understanding. In the example embodiment, the lid 20 and base 10 are each formed of a transparent material such that biological tissue stored therein may be visually observed throughout culturing and transportation.

[0039] FIG. 8 is a side profile view of an example carrier 200 (which may also be referred to as carrying container) and a plurality of stacked two-part containers 100 as previously described. In the example embodiment, the carrier 200 may be formed of a rigid frame 201 such as a wire like frame or skeleton. The frame 201 may include a first handle 202 and a second handle 203 that adjoin a support portion 204 for contacting a surface. Thespace between the handles 202 and 203 may form a receiving cavity 210 having a size and shape suitable for receiving a plurality of stacked containers 100 as described herein.

[0040] FIG. 9 is a graph showing the concentration of CCL21 protein in pg / ml at different time points during and after shipment for control and shipped thymus tissue cultures (PD = post day shipping). Culture Control: tissue slices left at 37 °C with daily media change. Sealed Control: tissue slices remain at ambient temperature for 48 hrs with no media changes. Shipped Cultures: tissue slices shipped at ambient temperature for 48 hrs.

[0041] Embodiments in accordance with the disclosure herein may be sterilized by various process such that the base 10 and lid 20 may be reused, e.g., by ethylene oxide (EtO) or by an E-beam sterilization method. Sterilization methods are known to degrade quality of containers and suitable material choices should take into account the sterilization processes and how that process may affect the durability of the O-ring 30 and the base 10 and lid 20. An example suitable material for forming the base 10 and lid 20 may be polystyrene. This material has shown to uphold well to e-beam doses of around 42 kGy (kilogray, an SI unit of absorbed radiation) such that there were no visual differences in color or clarity, and there were no cracks or deformities either. The O-ring 30 also upheld well at this irradiation level, and it should be appreciated that O-ring 30 can be replaced over time if and when necessary. Testing has shown that a suitable range of irradiation may vary from about 22-42 kGy.

[0042] Disclosed container 100 embodiments may be packaged together with other components to form a “kit” for sale and ease of use by various medical professionals. For example, disclosed herein is a kit type apparatus which may be used in a method of generating cultured thymus tissue, or portions thereof. In some embodiments the thymus tissue placed in the base of Container 100 is sourced from a donor. In some embodiments the thymus tissue is sourced from a recipient. In some embodiments the thymus tissue is cultured in the container 100 base. In some embodiments culturing is carried out by placing a conventional petri dish lid on the base of container 100 in lieu of the lid 20 shown in the detailed drawings. In some embodiments, following culturing, the screw top lid 20 is secured to the container 100 base and the thymus tissue is stored in the container 100 following culturing. Also disclosed herein is a kit for use in a method of generating a cryopreserved thymus tissue or a portion thereof. Also disclosed herein is a kit for use in a method of generating cultured thymus slices. Also disclosed herein is a kit for use in a method of generating cryopreserved thymus tissue slices, which may be cryopreserved before or after culturing. Also disclosed herein is a kit for use in a method of promoting donor specific tolerance in a recipient. Also disclosed herein is a kit for use in a method of providingimmunocompetence in a recipient. Also disclosed herein is a kit for use in a disclosed method of transplanting one or more solid donor organs in a recipient. Also disclosed herein is a kit for use in a disclosed method of performing an organ transplant in a recipient. Also disclosed herein is a kit for use in a disclosed method of reducing the risk of rejection of one or more solid organ transplants in a recipient. Also disclosed herein is a kit for use in a disclosed method of improving the viability of one or more transplanted organs in a recipient.

[0043] In an aspect, a disclosed kit as an apparatus and / or system can comprise any combination of reagents, compositions, tools, instruments, for coordinated use with and / or that are specifically adapted for use with disclosed container 100 embodiments which, as previously explained, may be used for culturing, procuring, storing, and transporting a thymus or a portion thereof. Such container 100 embodiments may also be useful for cry opreserving a thymus or portion thereof, thawing a cryopreserved thymus or portion thereof, slicing a thawed thymus or a portion thereof, maintaining thawed thymus slices, assaying thymus slices, implanting thymus slices, biopsying implanted thymus slices, or any combination thereof. In at least embodiment, a reagent may be an electron coupling reagent. In another aspect, the viability of the biological tissue may be assessed before transportation and / or upon receipt of the biological tissue at the subsequent transfer location. For example, assessing the viability of one or more prepared thymus slices to identify a thymus tissue sample or portion thereof prepared for implantation can comprise employing an MTS cell viability assay. MTS cell viability assays are known to the art. An MTS cell viability can comprise the quantitative ability of a thymus slice to bioreduce the MTS tetrazolium compound in the presence of an electron coupling reagent. The MTS cell viability assay can generate a viability index.

[0044] In another aspect, a disclosed kit as an apparatus and / or system may comprise any combination of reagents, compositions, tools, instruments, and container 100 embodiments for use in procuring a thymus or a portion thereof, slicing a thawed thymus or a portion thereof, maintaining thymus slices, cryopreserving thymus slices, thawing thymus slices, assaying thymus slices, implanting thymus slices, biopsying implanted thymus slices, or any combination thereof. In another aspect, a disclosed kit can comprise a plurality of components constituting the kit as an apparatus and / or system. Together, the plurality of components constitute a functional unit for a given purpose (such as, for example, preparing a thymus tissue sample or portion thereof for implantation). Individual member components may be physically packaged together or separately. For example, a kit comprising an instruction manual for using the kit may or may not physically include the instruction withother individual member components. Instead, the instruction manual can be supplied as a separate member component, either in a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as a recorded presentation or how to video.

[0045] In another aspect, a kit as an apparatus or system may be provided for use in a method that may include (i) one or more disclosed reagents, compositions, tools, instruments, sponges, filter paper, inspection tools, etc. and various container 100 embodiments, or any combination thereof, and (ii) a label or package insert with instructions for use. In an aspect, suitable supporting vessels may include, for example, bottles, vials, syringes, blister pack, etc. These supporting vessels may be formed from a variety of materials such as glass or plastic or any other suitable biocompatible material provided for in this disclosure. These supporting vessels may hold one or more reagents, compositions, tools, instruments, containers, or any combination thereof, and can have a sterile access port or packaging (for example the supporting vessels may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The label or package insert can indicate that one or more disclosed reagents, compositions, tools, instruments, container 100 embodiments, supporting vessels, or any combination thereof can be used in a method of culturing, preserving, protecting, transporting, and / or generating a thymus tissue sample or a portion thereof. Example methods may include generating a cryopreserved thymus tissue sample or a portion thereof, generating thymus tissue sample slices, promoting donor specific tolerance in a recipient, a disclosed method of providing immune-competence in a recipient, a disclosed method of transplanting one or more solid donor organs in a recipient, a disclosed method of performing an organ transplant in a recipient, a disclosed method of reducing the risk of rejection of one or more solid organ transplants in a recipient, a disclosed method of improving the viability of one or more transplanted organs in a recipient, or any combination thereof. Disclosed kits as an apparatus and / or system can comprise additional components necessary for administration such as, for example, other buffers, diluents, filters, needles, sterile water solution, and syringes.

[0046] In one example, a pre-packaged and sterilized kit of various components may be provided as a comprehensive packaged product. The kit may include a plurality of containers 100, a plurality of sterilized sponges for supporting biological tissue, sterilized water for maintaining appropriate humidity, buffers, reagents, culturing media, and syringes, and at least one carrier 200. The kit may further include a temperature logger, global positioning system trackers (gps trackers), instruction manual, scalpels, forceps, sterile gloves etc.EXAMPLE

[0047] An example custom container for culturing thymus tissue slices that maintains the integrity of the slices, allows for individual slice identity, and maintains an aseptic environment for the product was designed to transport thymus tissue slices to treatment centers. Twelve dishes in total were stacked in three stacks of four (as shown in Fig. 8), in a GasPak™ container (Becton, Dickinson & Co.), which was then placed in an example custom shipping container that was designed to maintain temperature control for the duration of the transportation. The shipping container was designed to be transported as carry-on baggage within a commercial airplane, and consists of insulation and phase change materials that ensure that the drug product remains within the specification of 15-25 °C for up to 96 hrs.

[0048] Dishes containing culturing media (Thymus Organ Media), and thymus tissue slices on filters (Millipore) that were placed on surgical sponges (Ethicon), were packaged for transportation via a commercial airline, from Raleigh, North Carolina to San Diego, California, followed by a return flight back to Raleigh, North Carolina. A courier accompanied the shipper throughout the duration of its transit. The thymus tissue slices were kept within the shipping containers for 48 hrs.

[0049] Pictures (not shown) were taken pre-shipment, during shipment, and at each end point, and show little to no movement of the slices during the transportation. Additionally, the slices were put back in culture for three days post shipment and the validated potency assay was performed to demonstrate that the activity and quality of the tissue slices was not impacted by the shipment (see FIG. 9).

[0050] The potency assay is an ELISA assay that allows us to measure the concentration of soluble, secreted CCL21, a signaling chemokine, in the culturing media (USVAL-02871 : Validation of Concentration Determination of CCL21 in thymus organ media samples by ELISA). The data in FIG. 9 supports that the shipment of thymus tissue slices in the custom dish is feasible and, in combination with the custom shipper, maintains the quality of the thymus tissue slices.

[0051] It should be understood that various aspects disclosed herein may be combined in different combinations than the combinations specifically presented in the description and accompanying drawings. For example, features, functionality, and components from one embodiment may be combined with another embodiment and vice versa unless the context clearly indicates otherwise. Similarly, features, functionality, and components may beomitted unless the context clearly indicates otherwise. It should also be understood that, depending on the example, certain acts or events of any of the processes or methods described herein may be performed in a different sequence, may be added, merged, or left out altogether (e.g., all described acts or events may not be necessary to carry out the techniques).

[0052] Unless otherwise specifically defined herein, all terms are to be given their broadest possible interpretation including meanings implied from the specification as well as meanings understood by those skilled in the art and / or as defined in dictionaries, treatises, etc. It must also be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless otherwise specified, and that the terms “comprises” and / or “comprising,” when used in this specification, specify the presence of stated features, elements, and / or components, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A two-part container for cultivating and / or transporting biological tissue, comprising: a lid having a first sidewall extending around a perimeter of the lid, the first sidewall including a first thread pattern; a base having a second sidewall extending around a perimeter of the base, the second sidewall including a second thread pattern corresponding in size and shape to the first thread pattern of the lid; and an O-ring disposed in an annular channel formed in the second sidewall; wherein, in a tightened position, the first sidewall of the lid compresses the O-ring against the second sidewall of the base thereby forming a protected storage space.

2. The two-part container of claim 1, wherein the base includes a floor having an interior surface for supporting biological tissue having a plurality of walls configured to partition the interior surface into a plurality of receiving areas.

3. The two-part container of claim 2, wherein the plurality of walls comprises at least eight walls configured to partition the interior surface into a first receiving area and a second receiving area.

4. The two-part container of any one of claims 1-3, wherein the lid and base each have a circular shape.

5. The two-part container of any one of claims 1-4, wherein the second sidewall of the base comprises a first flange and a second flange opposite the first flange portion.

6. The two-part container of claim 5, wherein each of the first flange and second flange extend a distance beneath an outside surface of the floor thereby forming a nesting cavity.

7. The two-part container of claim 6, wherein the nesting cavity defines a first area having a size, shape, and volume corresponding to a size, shape, and volume of the lid.

8. The two-part container of any one of claims 1-7, wherein the first sidewall comprises a plurality of ridges disposed around the perimeter of the lid.

9. The two-part container of claim 8, wherein the second sidewall comprises a plurality of ridges disposed around the perimeter of the base.

10. The two-part container of any one of claims 1-9, wherein the base includes a floor having an interior surface for supporting biological tissue and a plurality of labeled regions for identifying biological tissue.

11. The two-part container of any one of claims 1-10, wherein the base includes a floor having an interior surface for supporting biological tissue having a plurality of walls configured to partition the interior surface into a plurality of receiving areas, and wherein the plurality of walls are discontinuous such that moisture may be dispersed between the plurality of receiving areas.

12. The two-part container of any one of claims 1-11, wherein the base and lid are formed of polystyrene.

13. The two-part container of any one of claims 1-12, wherein the O-ring is formed of silicon.

14. The two-part container of any one of claims 1-13, wherein the base and lid are transparent.

15. The two-part container of any one of claims 1-14, wherein the O-ring is uncoated.

16. The two-part container of any one of claims 1-15, wherein, in the tightened position, the protected storage space defines a volume sufficient to hold enough oxygen to support biological tissue for at least 72 hours.

17. A method of using a two-part container for cultivating and / or transporting biological tissue, comprising: providing a two-part container, the two-part container including:(a) a lid having a first sidewall extending around a perimeter of the lid, the first sidewall including a first thread pattern;(b) a base having a second sidewall extending around a perimeter of the base, the second sidewall including a second thread pattern corresponding in size and shape to the first thread pattern of the lid; wherein the base includes a floor having an interior surface for supporting biological tissue having a plurality of walls configured to partition the interior surface into a plurality of receiving areas, and the plurality of walls are discontinuous such that moisture may be dispersed between the plurality of receiving areas; and(c) an O-ring disposed in an annular channel formed in the second sidewall; culturing a plurality of biological tissue samples in the base for about 5 days up to about 21 days, each biological tissue sample being disposed on a medical grade sponge in a respective receiving area; securing, after the culturing step, the lid to the base by engaging the first thread pattern and the second thread pattern thereby compressing the O-ring between the first sidewall and second sidewall to form a protected storage space, the storage space containing a sufficient volume of oxygen to support the viability of each biological tissue sample for at least 72 hours up to about 96 hours; and transporting the two-part container to a different location from where the culturing step took place.

18. The method of claim 17, further comprising: removing all of the biological tissue samples; sterilizing the two-part container by applying an e-beam irradiation technique comprising a dose of at least 22.5 kGy; repeating the culturing step with a new plurality of biological tissue samples; repeating the securing step; and repeating the transporting step.

19. The method of claim 17 or 18, wherein the plurality of biological tissue samples are thymus tissue samples cultured for about 19 days.

20. The method of claim 17 or 18, wherein the plurality of biological tissue samples are thymus tissue samples cultured for about 12-21 days.

21. A method of using, sterilizing, and stacking a plurality of two-part containers for cultivating and / or transporting biological tissue, comprising:providing a plurality of two-part containers, each two-part container including, respectively:(a) a lid having a first sidewall extending around a perimeter of the lid, the first sidewall including a first thread pattern;(b) a base having a second sidewall extending around a perimeter of the base, the second sidewall including a second thread pattern corresponding in size and shape to the first thread pattern of the lid and a first flange and a second flange defining a receiving cavity corresponding in size and shape to a circumference of each lid; wherein each base includes a floor having an interior surface for supporting biological tissue having a plurality of walls configured to partition the interior surface into a plurality of receiving areas, and the plurality of walls are discontinuous such that moisture may be dispersed between the plurality of receiving areas; and(c) an O-ring disposed in an annular channel formed in the second sidewall; sterilizing each two-part container via an irradiation technique; culturing a respective plurality of biological tissue samples in each respective base for about 5 days up to about 12 days, each respective biological tissue sample being disposed on a medical grade sponge in a respective receiving area; securing, after the culturing step, each lid to the corresponding base by engaging the first thread pattern and the second thread pattern thereby compressing the corresponding O-ring between the first sidewall and second sidewall to form a protected storage space, the protected storage space containing a sufficient volume of oxygen to support the viability of each biological tissue sample for at least 72 hours; stacking the plurality of two-part containers on top of one another; and transporting the plurality of two-part containers to a different location from where the culturing step took place.

22. The method of claim 21, wherein the plurality of biological tissue samples are thymus tissue samples that are cultured for about 19 days.

23. A kit apparatus for cultivating, storing, protecting, and / or transporting biological tissue, comprising: a plurality of two-part containers of any one of claims 1-16; a plurality of medical grade sponges for supporting biological tissue; and a carrier for stacking respective two-part containers.

24. The kit apparatus of claim 23, further comprising at least one reagent.

25. The kit apparatus of claim 23 or 24, further comprising sterilized water.

26. The kit apparatus of any one of claims 23-25, further comprising a temperature logger.

27. The kit apparatus of any one of claims 23-26, further comprising a GPS tracker.

28. The kit apparatus of any one of claims 23-27, further comprising at least one of: a syringe, sterile gloves, and a blister pack.

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