MXPA97009709A - Apparatus and method for the storage and transportation of bioartifical organs - Google Patents

Abstract

A bioartificial storage and transport apparatus having a media storage container, an assurance fixture and a gas exchange fixture and a media exchange fixture. In addition, the apparatus can use a secondary container and a lid, which has a gas access accessory and an access accessory to the media.

Description

APPARATUS AND METHOD FOR THE STORAGE AND TRANSPORT OF BIOARTIFICIAL ORGANS FIELD OF THE INVENTION The present invention relates to an apparatus and method for storing and transporting bioartificial organs ("BAO", for its acronym in English). The apparatus of this invention is designed to obtain BAO, so that the BAO is maintained under suitable culture conditions. This ensures the viability and sterility of the BAO until the implantation in the hall. Additionally, the apparatus of this invention is designed to minimize damage to the BAO during transport.

BACKGROUND OF THE INVENTION BAOs are devices that can be designed for implantation in a receiver or that can be developed to function extracorporeally. BAOs contain living cells or living tissue, which produce a biologically active molecule or provide a necessary biological function to an individual. In general, BAOs also contain a semipermeable membrane, which allows the diffusion of nutrients to cells and also allows segregated cellular products and waste materials to diffuse away from cells. In some cases, the membrane can also serve to immunoisolate cells by blocking the cellular and molecular effectors of immunological rejection. The use of immunoisolatory membranes allows the implantation of allogenic and xenogeneic cells in an individual without the use of immunosuppression. If the biologically active molecules are released from the isolated cells, they pass through the semipermeable membrane, surrounding to the body of the receiver. If metabolic functions are provided by the isolated cells, the substances to be metabolized enter the BAO from the recipient's body through the membrane to be activated by the cells. A variety of membrane types have been used in the construction of BAOs. In general, the membranes used in BAOs are membranes of either microporous or ultrafiltration grade. A variety of membrane materials have been suggested for use in BAOs, including PAN / PVC, polyurethanes, polysulfones, polyvinylidienes, and polystyrenes. Typical geometries of the membrane include flat sheets, which can be manufactured in "sandwich" constructions, having a layer of living cells placed between two essentially flat membranes with seals formed around the perimeter of the device. Alternatively, hollow fiber devices may be used, where living cells are located within a tubular membrane. The hollow fiber BAO can be formed gradually by loading the living cells into the lumen of the hollow fiber and by providing seals at the ends of the fiber. The hollow fiber BAO can also be formed by a co-extrusion process, where living cells are co-extruded with a polymer solution that forms a membrane around the cells. A common feature of the membranes useful in BAOs is that they are relatively thin walled and porous in nature. These membranes tend to be brittle and can be damaged during the manufacture and subsequent handling, transport and implantation of the BAO. BAO's have been described, for example, in U.S. Patent Nos. 4,892,538, 5,106,627, 5,156,844, 5,159,881, and 5,189,111, and PCT Applications Nos. PCT / US 94/07015 and WO 92/19195, all of which are incorporated herein by reference. in the present by reference. See also published PCT Applications Nos. WO 93/03901 and WO 91/00119. In addition to living cells and semipermeable membranes, BAOs may contain other components. For example, PCT publication No. WO 92/19195 discloses biocompatible vehicles as immunoisountable, implantable which have a hydrogel matrix to improve cell viability and which have a binding that aids in the recovery of the device. PCT Publication No. WO 91/00119 discloses membrane implants containing cells having external supports for connecting a plurality of tubular membrane-containing cells. PCT Application WO 93/21902 teaches implantable devices which have rigid and semi-rigid support structures. PCT Applications Nos. 94/07015 teach implantable devices which have holes to facilitate the introduction of cells and to provide airtight seals to cells, reliable. U.S. Patent No. 5,002,661 teaches a perfusion, pancreatic, intramuscular, artificial device having a housing for containing living cells dispersed in a matrix. BAOs that can be implanted can be inserted into a container in one of two ways. (1) implantation of a BAO semipermeable membrane device, vacuum followed by in situ addition of living cells, or (2) implantation of prefilled, cell-containing devices. In many cases, this latter approach is more desirable, due in part to the fact that the potential implant site may not be easily accessible for in situ filling. The performance of prefilled devices can also be characterized more easily in vitro. However, the storage and transport of the pre-filled devices present a number of problems. First, once manufactured, the BAOs must be maintained under adequate culture conditions to ensure the viability of the cells and the sterility of the devices until the time when the BAO is going to be implanted. In fact, in order to ensure that the BAOs are functional and sterile before implantation, the devices are maintained and tested over a period of many days. Sufficient evidence may require retention periods of approximately 17-24 days. However, longer retention periods than even a short period of time, such as about a week, require that the fluid media surrounding the BAO be periodically filled to provide a fresh source of nutrients and remove the waste products from the cells. encapsulated Additionally, the medium or fluid media will require sufficient dissolved gases, including oxygen, to maintain cell viability. It will be recognized that the nutrient and other requirements of BAO will depend on many factors, including the type of encapsulated cells, the metabolic activity of the cells, and the number and charge density of the cells. For example, chromaffin, adrenal, and bovine cells have been used in BAOs for implantation in human subjects for the treatment of chronic pain. Aebischer et al., "Transplantation in Hu ans of Encapsulated Xenogeneic Cells Without Immunosuppression," Transplantation, vol. 58, pp. 1275-77 (1994). It has been shown that these BAOs secrete various analgesic compounds, including norepinephrine, et-encafalon, octapeptide, and epinephrine. The BAOs are incubated in serum-free culture medium during the retention period. To maintain the release capacity of the analgesic substances, it is necessary to fill the medium after approximately 12 days. Typically, after filling, a three-day hold is made to ensure that the BAOs remain sterile. BAOs are cleaned for boarding after 15 days, and can be shipped until day 20. On day 20, the medium must be filled again, and a three-day retention is again required to verify sterility. Consequently, any storage system for the BAOs should allow easy access to the storage accessory to feed the BAOs. Another problem encountered with the use of pre-filled BAOs is that because they are not manufactured in general in the location where they are to be used, it is necessary to transport the BAOs to their implantation destination. For example, prior to the present invention, BAOs were frequently hand transported to the implantation destination in standard laboratory equipment, such as flasks or tubes with polypropylene screw caps. Since BAOs tend to be somewhat fragile, mainly due to the porous nature of the encapsulation membrane, occasional damage to the BAO has been observed during transit when embarking on normal laboratory equipment. Various packaging systems have been described for the storage and transport of body organs and tissue. See, for example, Tube, Published PCT Application No. WO 91/18575 ("Tube") and Krasner, Published PCT Application No. WO 91/03934 ("Krasner-1). Tube refers to a container for storage. and distribution of cultured epithelial skin wound cures The container comprises a pre-sterilized housing that supports wound healing in a medium of sodium bicarbonate, and a cover having a gas-tight seal to the housing. Gas is essential for container since the sodium bicarbonate medium requires 10% CO2 for the maintenance of the appropriate pH Krasner refers to a container for the storage and transport of bodily organs such as split limbs, or for storage and storage. conservation of bone graft material In Krasner, the bodily organs or the bone graft material are dropped into a mesh or gray hair that is suspended in a preservation solution reservoir. The bodily organs of the bone graft material are not secured in the container to prevent undue contact with the container and during transport. Bowman et al., US Patent No. 4,736,850, describes a device for collecting cells from a cut vein. The cell harvesting equipment includes a packing container that has three internal trays to support the components necessary for harvesting cells as well as the cut vein. The equipment is not designed to retain the fluid medium or to store the cut veins for anything other than a short period of time. In addition, the cut veins are not secured in the packing container to prevent undue contact with the container during transport. It is important that the exposure of the portions containing BAO cells to the air is carefully controlled. Prolonged exposure of BAO to the air will result in device drying that can be hazardous to cell viability. Preferably, the BAO is completely immersed in the fluid medium while it is secured in the storage and transport apparatus, regardless of the orientation of the apparatus. There is a need for an apparatus for storing and transporting the BAOs without substantially compromising the integrity, sterility and cell viability of the BAO.

SUMMARY OF THE INVENTION The present invention provides an apparatus and method for the storage and transport of BAO that substantially preserves the integrity, sterility and cell viability of BAO. The apparatus comprises a media storage vessel that bathes one or more BAOs in a suitable volume of the medium, a means for securing the BAOs within the media storage container, preferably such that the BAOs remain immersed in the liquid medium without consider the orientation of the BAO, and a means for sealing the media storage container designed to provide a substantial seal fluid-tight body. The securing attachment is designed to prevent damage to the BAO resulting from turbulence or undue physical contact with other elements of the media storage container. Where the BAO is going to stay in the fluid medium P5 > for more than a short period of time (i.e., about a week), the apparatus preferably has a medium exchange accessory and a gas exchange accessory to maintain the viability of the cells in the BAO. The fluid exchange fitting allows the provision of fresh media and the removal of waste products, while the gas exchange accessory allows the introduction of oxygen into the medium to keep the cells alive in the BAO. The apparatus also optionally comprises a secondary container that surrounds the media storage container. The use of a secondary container minimizes the handling of the media storage container, thereby reducing the risk that the sterility of the BAO may be compromised. The secondary container also facilitates the introduction of the bioartificial organ into a sterile field at the time of implantation.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows a preferred storage and transport apparatus of BAO. The apparatus comprises a container 10 for storing the medium or media and a lid 12. A secondary container 100 and a secondary lid 102 are also shown.

Figure 2 shows a cross-sectional view of a storage and transport apparatus of BAO comprising the storage container 10 of the medium and the lid 12, contained within the secondary container 100 and the secondary lid 102. Figure 3 shows a storage and transport apparatus of BAO comprising the storage container 10 of the medium and a alternative embodiment of the lid 12. Figure 4 shows a view of the upper part of the medium storage container 10 and the securing means 20 holding or holding a BAO. Figure 5 shows the effects of temperature and time on condensation inside the BAO storage and transport apparatus during shipping or shipping.

DETAILED DESCRIPTION OF THE INVENTION In order that this invention may be more fully understood, the following detailed description is set forth. The present invention provides an apparatus and method for storing and transporting BAO's. In general, the apparatus comprises a media storage container that retains or maintains a volume of fluid media, a means for securing the BAOs within the media storage container, and a means for sealing the media storage container designed to provide a seal substantially fluid-tight. Preferred embodiments further comprise a media exchange fitting and a gas exchange fitting to maintain the viability of the cells in the BAO. The apparatus also optionally comprises a secondary container that surrounds the media storage container. Preferably, the secondary container has a means for accessing the exchange accessory of the inner container (the media storage container). Additionally, the secondary container may have a means for exchanging moisture from the interior of the secondary container to the external atmosphere to prevent or prevent excessive accumulation of moisture within the secondary container. In a more particular way, the BAO storage and transport apparatus of this invention comprises a media storage container 10 and a lid 12. Typically, the media storage container 10 has a cavity 14 for facilitating media exchange. The cavity 14 can have any suitable design, including a groove formed around the bottom of the media storage container 10 and / or a low point on the bottom. The presence of this cavity or low point provides a channel for directing air bubbles away from the BAO, if the media storage container 10 is inadvertently transported or stored in an upside down orientation. Preferably, the media collection cavity 14 should be located away from the semi-permeable membrane of the BAO, brittle to ensure that the BAO is not damaged during any of the media exchange procedures. In one embodiment, the media storage container 10 has a convex base 16. In addition, the circumference of the base 16 forms the cavity or groove 14 around the bottom of the media storage container 10. An securing means 20 for securing the BAO inside the media storage container 10 can be attached to the interior of the media storage container 10 or the bottom of the lid 12. The securing accessory can also comprise an insert which is fits into the media storage container 10. Preferably, the securing means 20 is attached to the base 16. Under normal operating conditions, the securing attachment 20 Pt > 2 b keeps the BAO immersed in the fluid media, so that the BAO is not subjected to undue contact with any component of the system other than the securing accessory. Any suitable securing accessory can be used to secure the BAO. The person skilled in the art will appreciate that the exact design of the securing fitting 20 will vary according to the design of the BAO. As discussed above, BAOs are devices that contain living cells or tissues encapsulated in a semipermeable membrane. The membrane can be sealed in a variety of ways, such as with caps, glue or thermal seal, to form either a cell compartment or multiple compartments. In addition, BAOs may also have a means to recover the device after implantation, such as a tie attached to a sealed end. Preferably, the securing means 20 will not physically compress the portions of the semi-permeable BAO membrane. It is contemplated that the securing fitting contacts the BAO in a non-membrane portion of the BAO, such as tie, brackets, seals or connectors. The securing means 20 must provide a substantially unrestricted flow of the fluid media over the BAO, which permits the free exchange of nutrients and waste products. Preferably, the securing means 20 allows the BAO to be floating freely in the media. In a preferred embodiment shown in Figures 1 and 2, the securing attachment comprises teeth 20 and the post 22. As shown, the teeth 20 and the post 22 are attached to the base 16 of the media storage container 10. It is contemplated that the teeth 20 and the post 22 can be placed in the base 16 to accommodate the BAO of various sizes. In the preferred embodiment, the teeth 20 and the post 22 are designed to attach a BAO capped at one end with a fitting sealed in the center and having a tie attached to the lid. Sealed BAOs at the center are taught in PCT / US / 94/07015. The BAO sealed in the center is secured by placing the tie on the tooth 20 such that the non-membrane portion of the BAO (eg, the lid sealed at the center) is equated with the outer surface of the tooth 20, and then rolled up the tie around the post 22. The BAO is thus secured at one end only, with the semi-permeable and fragile portion floating freely in the central portion of the media storage container 10. The BAO can be additionally secured to retain the tooth 20 and / or the post 22 P525 by the use of appropriate tweezers or slots, or other suitable means. It is also contemplated that the teeth 20 can couple and secure the BAOs of other designs. For example, BAOs having non-membrane, external, features (eg, supports for connecting a plurality of membranes containing cells) can be secured within the media by joining the teeth 20 around the non-membrane portion. In addition, the assurance accessory can be designed to ensure a BAO in more than one location. This can be achieved by using any combination of teeth 20 and / or posts 22 as long as the integrity of the functional membrane portion of the BAO is not compromised. For example, where the BAO has non-membrane portions at both ends of the BAO (e.g., end supports and seal caps as described in Published PCT Application No. WO 91/00119), the teeth 20 can be aligned so that each non-membrane portion can be inserted through the slot of a different tooth 20. The lid 12 provides a substantially fluid-tight seal when it is seal-coupled with the media storage container 10. In the preferred embodiment, when the lid 12 is engaged, the media storage container 10 is invertible so that the BAO T525 remains submerged in the middle and does not come into contact with the air bubbles that will tend to accumulate in the cavity or low point of media collection. This ensures that during transportationIf the container is inadvertently inverted, the BAO does not dry out. Preferably, the cap has a liner (e.g., a compressible material such as silicone elastomer) that aids in the formation of a fluid tight seal. Additionally, the lid can be designed so that a portion thereof extends into the cavity of the means, thereby causing an attachment of the media to be ejected from the container. This serves to minimize the formation of air pockets within the storage and transport apparatus filled with media when sealed. This ensures that the BAO is completely enclosed by fluid media during storage and transport. The liner can be formed with a layered construction approach in which a layer of silicone is placed between two layers of other suitable polymers (eg, polypropylene or fluoroethylenepropylene). The liner can be fixed by any suitable means, such as ultrasonic welding. Where the lining is formed by layered construction, the inner layer of the liner, which makes contact with the lid, will be made of the same material as the lid in order to facilitate ultrasonic welding. The outer layer can be of any suitable material that will be low friction and durable enough to form a fluid tight seal. For example, fluorinated perfluoroethylene-polypropylene (FEP), a copolymer of hexafluoropropylene and tetrafluoroethylene, is a suitable material for the low friction layer of the liner. FEP has very similar properties such as polytetrafluoroethylene (PTFE), but is more durable during sterilization with gamma irradiation than PTFE. Where a liner is used, it is preferred that the media storage container 10 have a sealing edge around its outer perimeter, so that when the lid 12 is attached to the container, the sealing edge will fit into the compressible liner for form the hermetic seal to fluids. One embodiment of the lid 12 is a lid which is heat sealed to the upper part of the media storage container 10. The thermal seal equipment is well known. Where a liner is used, it may be comprised of bonded olefin by spinning, paper, aluminum foil, or similar materials known in the art. In this embodiment, the liner contains a resin that attaches the liner to the container 10 when heat is applied. In another embodiment, the lid 12 is a threaded lid that engages the connecting threads in the upper part of the media storage container 10. In a further embodiment shown in Figure 3, the lid 12 is a threaded lid having a recessed region that ejects the media from the media storage container 10 as the lid engages the joint threads on the top of the container. This minimizes the formation of air pockets within the media container when it is sealed, and furthermore ensures that the BAO is completely surrounded by the fluid media during storage and transportation. A preferred embodiment of the storage and transport apparatus shown in Figures 1 and 2 further includes a gas exchange accessory 50 and a media exchange accessory 30. The ability to exchange gas to the media aids in the maintenance of viability in living cells within the BAO. Alternatively, sufficient oxygen can be introduced into the packing system by saturating the media before attaching the sealing fittings, or by using a breathable liner. As shown, the gas exchange fitting 50 is a resealable hole that allows gaseous communication between the outside of a media storage container, sealed therein. He P525 orifice can be resealed by any suitable method, such as a cap, stopper, or preferentially a self-sealing septum 52. These septa are well known in the art. In embodiments using a self-sealing septum, to permit gas exchange, a suitable hollow needle or alternative is inserted through the septum 52 into the media in the media storage container 10. The needle preferably contains a microbial filter (for example, 0.22 micron PTFE filter in a polypropylene housing) to avoid contamination of the media storage container 10, and thus, the BAO, with contaminants in the introduced gas . Preferably, the gas exchange fitting 50 also includes a stop member for limiting the extent to which a needle can be inserted into the media storage container 10. The stop member prevents the needle from coming into contact with the BAO, and thus prevents accidental drilling of the BAO. The media exchange accessory 30 is similar in design to the gas exchange accessory 50. For example, the media exchange accessory 30 may also comprise a re-sealable hole for the I ",". access to the media. Preferably, the media exchange accessory 30 has access to the media collection cavity 14. The media can be removed from or inserted into the media storage container 10 using a needle, tube, or other suitable methods. Preferably, the media exchange attachment 30 includes the self-sealing septum 32, which is directly aligned over the media collection cavity 14. To exchange the media, a needle is inserted through the septum 32 such that the opening of the needle is placed in the cavity. The media in the cavity is then aspirated and removed from the system. Fresh media can be introduced in a similar way. Preferably during media exchange, the media passes through a suitable filter to prevent the introduction of some contaminants into the media storage container 10. After the media exchange occurs, the needle is removed and the septum 32 self-packs to form a fluid-tight seal. Where the gas exchange fitting 50 and the media exchange fitting 30 use the self-sealing septa, a preferred embodiment uses quick attachments to accommodate the septa. For example, base 58 of gas exchange rapid adjustment and base 30 of quick adjustment of media exchange are inserted through the bottom of the appropriate holes in cover 12 such that a portion of the bases 58 and 38 of adjustment Rapid extends through the top of the lid 12. The septa 52 and 32 are then inserted into the quick-fit bases 58 and 38. The containment rings 54 and 34 fit comfortably around the portion of the quick fit bases 58 and 58 that extend through the top of the cover 12. The quick fit inserts 56 and 36 then join together. to the upper parts of the quick adjusting bases 58 and 38, respectively. The quick-fit bases and the quick-fit inserts are designed to pull each other when they are joined, thereby ensuring a fluid-tight seal around the septa 52 and 32 and the liner of the lid. The quick-fit housings for either or both of the gas exchange fitting 50 or the media exchange fitting 30 can also serve as a handle for the media storage container 10. As shown in Figure 1, the quick-fit insert 56 works like a handle. The components for these accessories can be manufactured by machining, injection molding, or other common techniques. As an alternative to the quick fittings, inserts can be ultrasonically welded onto the cover 12.

In another embodiment, means may be provided by one or more additional containers, integral to the media storage container 10. In this embodiment, the spent media is drained from the media storage container 10 via the pipe connecting the media storage container 10 to a spent media container. The fresh media is introduced from a reservoir of fresh medium in fluid connection with the media storage container 10. More preferably, the media exchange reservoirs can be connected from the media storage container 10 without compromising the sterility of their contents. Once disconnected, the reservoirs can be drained or replenished, as appropriate, and then reconnected to the media storage container 10 in a sterile manner. These procedures are known in the art. Figures 1 and 2 further show the secondary container 100 and the secondary cover 102, which enclose the media storage container 10 and the lid 12. The secondary cover 102 provides a substantially fluid-tight seal when it is sealed with the container Secondary 100. In one embodiment, the secondary cover 102 is a threaded cover that engages the connecting threads in the upper part of the secondary container 100. Preferably, the secondary cover 102 has a liner, as described above for the cover 12 In addition, the secondary lid 102 preferably has a means for access to the gas exchange fitting 50 and the media exchange fitting 30 as well as a means for releasing moisture. These access fittings and the moisture exchange fitting are similar in design to the exchange means 50 and 30 in the lid 12 of the media storage container 10, but may have any other suitable design. Shown in Figure 2 is a preferred embodiment having an access fitting 150 for the gas and the accessory 130 for accessing the media. As shown, the access fittings 150 and 130 have self-sealing septums 152 and 132 in quick-fit housings (ie, the bases 158 and 138, which contain the rings 154 and 134, the inserts of the top 156 and 136). The gas access fitting 150 is aligned with the gas exchange fitting 50 and the media access fitting 130 is aligned with the media exchange fitting 30, such that a needle or other suitable instrument can be inserted. , from the external environment through the septa of the secondary container and then through the septa of the media storage container, thereby penetrating the media storage environment to allow the exchange of gas and / or media. In a preferred embodiment, the bottom of the media storage container 10 has a key mechanism 60, which engages in a bonding base 200. A key mechanism 60 ensures that the access means 150 and 130 of the secondary lid 102 are aligned over the exchange accessories of the lid 12. In addition, the key mechanism 60 allows an operator to align the media exchange attachment 30 of the lid 12, with the media collection cavity 14 of the base of the media storage container 10. This is achieved by png the media storage container 10 on a base plate that is similar in design to the junction base 200. Additionally, where the lid 12 is a threaded lid, the key mechanism 60 also facilitates the joining or detachment of the lid 12 without excessive handling of the media storage container 10. The key mechanism 60 allows an operator to secure or remove the lid 12 simply by adjusting or fitting the media storage container 10 to a joint base similar to the joint base 200 and then applying a torque or torque to the lid 12. The reduced handling of the media storage container 10 is desirable due to the reduced contamination potential. In a preferred embodiment, shown in Figures 1 and 2, the storage and transport apparatus further includes the moisture exchange accessory 170 which is similar in design to the gas exchange accessory 50. The moisture exchange attachment is placed in the secondary cap 102 and comprises the septum 172 having a hole in which a breathable liner 179 is placed. The breathable lining 179 may be, for example, a membrane disk with a porosity of 0.22 microns of nylon. A nylon membrane is preferred since it is capable of avoiding sterilization procedures, such as gamma irradiation. A porosity of 122 microns is preferred to prevent microbial contaminants from entering the interior of the secondary container. The moisture exchange attachment 170 allows moisture release and pressure equalization during transport to prevent condensation from forming inside the secondary container. The exchange of moisture is desirable since the container can be stored in a humidified atmosphere incubator. Without the moisture exchange attachment 170, moisture can be started and remain inside the secondary container as the temperature drops from P5 'approximately 37 is in the incubator until room temperature during boarding. In Figure 5, the effect of the loss of temperature in the condensation is graphically represented. Each of the three runs shown was carried out in a storage and transport apparatus of this invention in which the secondary container included the moisture exchange attachment comprising a membrane disk of porosity of 0.22 microns of nylon placed in the Secondary cap inside a septum that has a hole with an inside diameter of 0.312 inches. The initial temperature of the apparatus for each run was approximately 37 se. Run-1 represents conditions of "rapid cooling" in which the temperature of the apparatus was dropped rapidly from about 37 ° C to about 25 seconds in one hour. Under these conditions, condensation appears, but over time, the interior of the secondary container will dry out as the condensed moisture evaporates through the membrane disc. Run-2 represents "slow cooling" conditions in which the temperature of the apparatus is controlled. Under these conditions; the membrane disc seems to have prevented condensation from forming inside !: "- >. ' of the secondary vessel during the test period, Corrida-3 represents simulated operating conditions (actual thermal and time conditions expected during a routine commercial shipment) During this run, the vessel was held to the shipping or shipping conditions. simulated during a period of approximately 8 hours, after which time the container was exposed to "rapid cooling" conditions.The condensation did not appear visibly during the simulated loading period.As with the "slow cooling" run, the disc The membrane seems to have prevented condensation from forming inside the secondary container Only after the apparatus was exposed to the conditions of "rapid cooling", condensation appeared The quick-fit housings for the gas access accessory 150, the media access accessory 130 and the moisture exchange accessory 170 can be constructed in a similar manner the gas exchange accessory 50 and the media exchange accessory 30. The quick-fit bases 158, 138 and 178 are inserted through the bottom of the appropriate holes in the secondary lid 102. The septa 152, 132 and 172 are then inserted into the quick-fit bases 158, 138 and 178, respectively. With regard to accessory 170 of P5? Moisture exchange, septum 172 has a hole in which the breathable lining 179 is inserted. The containment rings 154, 134 and 174 are fixed around the quick-fitting bases 158, 138 and 178, respectively, to secure the quick fittings to the secondary caps 102. Finally, the insertion pieces 156, 136 and 176 fast are attached to the top of the bases 158, 138 and 178, of fast stment, respectively. It is preferred that the quick-fit insert 156 include a needle-holder array while the snap-fit insert 136 provides a septa cover 139. Alternatively, as with the exchange accessories 30 and 50 of the caps 12, the quick fittings used with the access accessories 130 and 150 as well as with the moisture exchange accessory 170, may be replaced with inserts that are ultrasonically welded on the cover 102. The secondary container 100 minimizes the need to handle the main container 10 once the BAO and media are initially loaded. Subsequent exchanges of gas and media can be carried out through both containers with minimal disturbance of the storage and transport apparatus and with a substantially reduced risk of contamination of the container.

Pr) 2í > BEAM. In the distribution to the implantation destination the secondary container 100 can be opened out of the sterile field, the media storage container 10 can be distributed to the sterile field, thereby reducing the potential for the introduction of unwanted biological contaminants into the sterile field. If the secondary container 100 is used, the grooved joint base 200 may take the form of a disc attached within the base of the secondary container 100, for example, by ultrasonic welding. The grooved joint base 200 has several clamping tongues 202 along its upper edge. These fastening tabs 202 are joined with the key mechanism 60, which are shown in Figures 1 and 2 as a series of slots in the bottom of the media storage container 10. When the media storage container 10 is inserted into the secondary container 100, the slots receive the securing tabs 202. Alternatively, the secondary container and the grooved joint base can be manufactured as a single unit, using for example injection molding. . The media storage container 10 should fit comfortably into the secondary container 100 in order to prevent movement of the media storage container which could result in damage to the BAO. This adjustment or quick accommodation can be easily achieved by the proper design of the height of the gas exchange fitting 50. Any suitable material can be used in the manufacture of the storage and transport apparatus, including metal, such as stainless steel, aluminum alloys or titanium based materials, or ceramic products, such as silica or alumina-based ceramic products, available for example from Coors Company, Boulder, Colorado. Any material that is capable of withstanding common sterilization methods can be used, such as irradiation (i.e., gamma irradiation), chemical methods (i.e., sterilization with ethylene oxide), and autoclaving. If the sterilization method is gamma irradiation, the chosen materials, if necessary, should contain stabilizers to the gamma radiation to prevent discoloration or deterioration. Commercially available polycarbonate polymers have these stabilizers. These polymers are typically colored (eg, stained purple) before gamma stabilization. In exposure to gamma radiation, the polymers turn a yellowish color; however, if they are properly stabilized to gamma irradiation, these colored polycarbonate polymers can be reversed back to their natural dye after a cut or period of reasoning. In addition, the materials must be durable enough to withstand the rigors of boarding and handling. Additionally, the materials must be sufficiently non-toxic to the living cells so as not to produce appreciable harmful effects to the BAO. Preferably, the materials must be tested with the USP class 6. While the reusable materials are contemplated, preferentially, the materials are disposable. It is also preferred that at least some of the components of the storage and transport apparatus be optically transparent. This will allow the easy discovery of visible contamination of the BAO. Examples of visible contamination include turbidity and discoloration of the fluid media. Preferably, the media storage container 10 and the secondary container 100 are constructed of optically transparent materials such as glass, or a suitable polymer. Useful polymers include any non-brittle, durable material such as polycarbonate, polyphenylene oxide, polyesters, polyetheramide, polyethylene terephthalate, terephthalate Pb? polyethylene co-glycol, and similar resins. Polycarbonate polymers are most preferred because: (1) they are more durable than some other suitable materials, (2) they are available in class 6 grades of USP, and (3) they are thermoformable. The containers and caps of this invention can be made by any suitable technique. For example, they can be constructed by injection molding or other thermoforming methods. Alternatively, they can be machined, or assembled using joining techniques such as ultrasonic welding. The preferred method of manufacture is a thermoforming process such as injection molding, since this process easily provides for the production of complex shapes. Containment rings that are produced from polycarbonate materials by machining or injection molding have stress lines that are amplified by exposure to gamma irradiation during sterilization. It is desirable to remove the lines of effort in order to ensure the hardness of the plastic. Tempering can be done to reduce stress imbalance. Exposing the rings machined or molded at a temperature of approximately 2502C for approximately 10 minutes and then slow cooling (for several hours) to prevent the P525 thermal shock, accelerates the stress relaxation process that occurs in all thermoplastics. When tempering, the internal efforts are reduced which improves the dimensional stability and the performance of the parts. When devices are shipped, care must be taken to minimize the physical rigors of transportation. As a result, a tertiary vessel can be used for shipping or shipping to enclose the storage and transportation apparatus. In addition, it may be desirable to control and inspect the BAO temperature during transport. The tertiary shipping container can be packaged with insulating, thermal, or similar gel packs, such as are commercially available from Packaging Products, Inc. of New Bedford, MA. The shipping or shipping containers themselves may be insulated, and it may be desirable to include an inversion indicator or temperature recorders to ensure proper handling of the BAO temperature. Isolated containers with reversal indicators are commercially available (eg, the isolated EnduroTherm vessel, from Source Packaging of New England; Warwick, IR). Any suitable temperature zone can be used (e.g., Data Trace, Mesa Laboratories, Inc., Wheat Ridge, CO).

The shipping or shipping container must be durable and capable of absorbing the blow when dropped or other movement during boarding to protect the BAO from damage. Insulated shipping containers that have thick layers of insulation (1-3 inches) are useful for absorbing bumps from the container during shipping. The invention is further illustrated by the following example, which is not to be seen as limiting in any way.

EXAMPLE The apparatus for storing and transporting a BAO is assembled and comprised of a media storage that retains approximately 80 mol of a suitable fluid medium. The media storage container has a cavity, or low point, positioned away from the center of the base of the media storage container. The media storage container was made from an optically transparent polycarbonate material and formed by injection molding, using a machine-made mold of stainless steel or aluminum. The media storage container had an outer diameter of approximately 3.78 inches and a sealing edge of 0.1925 inches in width. The inside diameter of the media storage container was approximately 3.40 inches and could be filled with approximately 80-100 ml of the medium to immerse the BAO. The threading of the storage container was of an SP-400 finish as described by the Plástic Bottle Institute, Technical Committee, a division of the Society of Plastics Industry Incorporated. Additionally, the identification of the thread termination was of size 100 and preferably a modified reinforcing thread. The securing attachment was comprised of a number of posts and teeth positioned perpendicular to the bottom surface of the media storage container. The height of the posts was such that the tops of the posts would extend to the bottom of the sealing fitting. This additionally prevented the BAO from undue movement within the storage container. Poles and teeth are an integral part of the base and are formed by molding or injection. The teeth have a tapered, slotted section about 0.026 inches wide in order to tighten the BAO tie section that was 0.030 inches in diameter as it slid into place. The polycarbonate material from which the poles and teeth are made, allowed the tie to be held firmly and was hard enough not to break under the load.

P525 Poles and teeth were placed along the bottom of the container such that the media container could safely hold a BAO of active length of 5 cm and / or an active length of 7 cm such that the membrane portion containing BAO cells would be in the center of the media storage container. The BAOs used in this example have hollow, compressible silicone bonds attached to one end of the BAO with an outer diameter of approximately 0.030 inches. The sealing fitting used was in the form of a threaded cover which engaged the attachment threads on the bottom at the top of the media storage container. The cover thread was a modified 5 TPI, # 100-400 support thread as described by the specifications of the Closure Manufacturers Association. The lid was coated with an elastomeric, compressible silicone material to aid in the formation of a fluid tight seal. This liner was made from a layered construction by co-current silicone description methods between a layer of a polypropylene film and an FEP layer. The silicone was 0.040-0.100 inches thick and the films were 0.005-0.10 inches thick. The liner was fixed to the lid by ultrasonic welding. The inner layer of the liner was made from the same material P525 as the cover, specifically polypropylene, to facilitate ultrasonic welding. The ultrasonic welding of the lining was achieved by the energy of direct bonding to the materials. Six welding spots, each 0.375 inches in diameter, were symmetrically applied around the lid in order to firmly attach the liner in place. The media storage container tubes a 902 blade edge approximately 0.010 inches high around the outer edge, which cut into the compressible silicone and FEP portions of the lid liner to aid in the formation of a fluid tight seal . The bottom of the media container has a key mechanism that fits into a joint base and / or a base plate. The key mechanism, a misaligned tongue ios, allowed the alignment of the pierceable septum over the cavity of the media and also for the alignment of the septa of the secondary container over the septa of the media storage container. The apparatus of this example contained both a media exchange accessory and a gas exchange accessory. These exchange accessories included self-sealing silicone septums. The media was exchanged when a needle was inserted through a silicone septum approximately 0.125 inches thick and 0.550 inches in diameter, followed by aspiration and removal of media. Fresh media was introduced with another appropriate needle through the same silicone septum. A lid attachment was used to cover the septa during storage. Gas exchange was performed by inserting a hollow needle through the second septum. The needle contained a 0.22 micron Teflon filter in a polypropylene housing held in the needle with a leur fitting. The septum in a gas exchange additionally comprised a stop member which limited the extent to which the needle could travel inside the media storage container. For long-term storage, the device is placed in storage incubators with the needle inserted through the septum. If the device is to be shipped, the needle is removed during the brief shipping or shipping period and the septum self-packs to provide a fluid-tight seal. The materials used for the manufacture of the apparatus were polycarbonate, polypropylene and silicone and are completely disposable, and were not reused. These materials were selected as they can withstand gamma sterilization methods, sufficiently durable to withstand the rigors of shipping or shipping and are sufficiently non-toxic to living cells to not produce appreciable detrimental effects to BAOs. The materials were all tested in class 6 of the USP. The apparatus further comprised a secondary container that encircled the media storage container, sealed. The secondary container tube an access fitting to the gas and media, both of which were self-sealing septums aligned on the self-sealing septa of the media storage container. In this example, the secondary containers similar to the media storage container since it had a similar cover construction, similar to self-sealing septa for access to the interior of the media storage container. The secondary container also had a half moisture exchange of a 0.22 micron nylon membrane having a diameter of 0.312 inches. Access to the interior of the media storage container was achieved by aligning the septa of the secondary container on the septa of the media storage container such that a needle can be inserted through the septa into the secondary container and through the septum into the septum. primary vessel to provide the exchange of media or gas. The apparatus used in this example was assembled (uncoupled threaded lids) that include liners P525 cover, septa, quick fittings, and the base ring of the secondary container and placed in., Bags for sterilization by gamma irradiation. The sterile units were then placed in a sterile hood where the BAOs were loaded.

BAO Loading Procedures BAO charging is performed according to the following procedures: (1) the media storage container was placed on the base plate such that the union tabs on the base plate were fitted into the slots at the bottom of the media storage container; (2) 10 ml of the selected media were added to the media storage container in order to wet the bottom of the container; (3) the BAOs were then loaded, as shown in Figure 4, by placing the thin, cylindrical BAOs in the center of the media storage container; (4) the silicone tie was inserted into the teeth and then wound around the posts; (5) the media storage container was then hermetically sealed with the threaded cover using the base plate to retain the container so that the knife edge was cut into the cover liner; (6) the alignment of the media exchange fitting on the media cavity of the media storage container was then visibly confirmed; (7) The media storage container sealed with the BAO inside was then placed in the secondary container on top of the grooved joint base, such that the tabs of the joint base were fitted into the slots of the key mechanism at the bottom of the media storage container; (8) the secondary container was then hermetically sealed with the threaded cover so that the knife edge cover was cut into the liner of the cover; (9) the gas exchange needle and filter housing of 22 microns were then placed through the septum of access to the secondary container gas and then through the gas exchange septum of the media storage container; (10) The media exchange needle was then placed through the secondary container gas access septum and then through the gas exchange septum of the media storage container, such that the pointed needle entered the cavity of the media; and (11) the remaining 70 ml of the media were then added through the media exchange needle, thereby filling the media storage container.

Transportation and discharge procedures. The BAO-containing apparatus, loaded then, was distributed to the implantation destination according to the following procedures: (1) the gas exchange needle was removed from the media storage container; (2) the apparatus was placed in a tertiary, insulated and durable packaging box that has temperature inspection devices and thermal packaging; (3) the apparatus is then shipped or shipped to the destination of implantation via routine transportation methods; (4) At the implantation destination the apparatus was removed from the shipping package and the lid of the secondary container was uncoupled so that the sterile, loaded and sealed media storage container could be introduced into the sterile surgical field (5) once inside the sterile field, the lid was uncoupled in the media storage container; and + (6) the BAO then removed the media storage container by unwinding the BAO tie from the posts and uncoupling the fastening of the teeth.

Test procedures before implantation Prior to implantation in patients, BAOs should generally be functional and sterile. This test may require retention periods of approximately 17-24 days for termination. During this period of time, it may be necessary to change the means to maintain cell viability. For example, as described above, where BAOs contain bovine adrenal chromaffin cells, it is necessary to fill the medium after approximately 12 days. After the media exchange, a short three-day retention is performed to ensure the sterility of the BAO. BAOs then embark after day 15, if not until day 20. On day 20, the cells should be fed again after another short retention of three days.

Product evaluation procedures 1. Sterility test The BAOs were loaded and media exchanged as described above and the BAOs were stored in the apparatus for 29 days. Sterility was assessed through the use of USP sterility evaluations, the devices were found to remain sterile.

P525 2. Impermeability test The media storage container, seal and sealed secondary container were evaluated for air-tight and fluid seals. The air tightness test was performed by the addition of 1-3 psi of compressed nitrogen, and was evaluated for pressure maintenance over time. The devices were found to remain air tight at a pressure of 1-3 psi for extended periods of time. The fluid-tight seal was assessed by adding media, inverting the containers, and rotating the media inside the containers for extended periods of time. The devices were found to remain fluid-tight. In addition, the devices were shipped unattended on the Island of Rhode by a common carrier to a site in Virginia. The devices were made and there was no evidence of violation of the liquid tight seal. The drop test from the vertical of 10-20 feet was also performed in order to ensure that no damage would occur during transport. (3) Toxicity test. The toxicity of the manufacturing materials was valued when evaluating the BAO production for analgesic substances after being stored for 29 days, P525 including the repositioning of the media on days 12 and 20 as described above. In addition, the cells used in the BAO were placed directly in the materials and evaluated in a similar manner for the production of active substances and changes in cell morphology. It was found that the materials are non-toxic neither to the cells nor to the BAO. The analgesic substances were not changed by the exposure of the materials.

P.r) 2

Claims (25)

1. CLAIMS: 1. A device for storing and transporting bioartificial organs, comprising: a media storage container having a securing accessory attached thereto for securing the bioartificial organ to the media storage container; a lid that joins the media storage container to form a fluid tight seal; where the bioartificial organ is immersed in the fluid media. The apparatus according to claim 1, wherein the media storage container is made from an optically transparent material. The apparatus according to claim 2, wherein the media storage container is made from polycarbonate polymers. The apparatus according to claim 1, wherein the securing attachment comprises at least one retaining tooth and at least one post. The apparatus according to claim 1, wherein the cap comprises a threaded cap, and the media storage container has connecting threads that engage the threaded cap. P 2 S 6. The apparatus according to claim 1, further comprising a gas exchange fitting. The apparatus according to claim 1, further comprising a media exchange accessory. The apparatus according to claim 1, further comprising a gas exchange fitting and a media exchange accessory. The apparatus according to claim 9 or claim 8, wherein the gas exchange fitting comprises a resealable septum. The apparatus according to claim 6 or claim 8, wherein the gas exchange fitting provides the sterile, gas exchange between the media storage container and the external medium. The apparatus according to claim 7 or claim 8, wherein the media exchange accessory comprises a resealable septum. The apparatus according to claim 7 or claim 8, wherein the media exchange accessory provides the sterile media exchange between the media storage container and the external media. 13. The apparatus according to claim 1, which P525 further comprises: (a) a secondary container capable of enclosing the media storage container when it is attached to the lid; and (b) a secondary lid that attaches to the secondary container. The apparatus according to claim 13, wherein the secondary container or the secondary lid further comprises a moisture exchange accessory. The apparatus according to claim 6, further comprising: (a) a secondary container capable of enclosing the media storage container when coupled to the lid; and (b) a secondary lid that attaches to the secondary container. The apparatus according to claim 15, wherein the secondary container or the secondary cover further comprises a gas accessing accessory for accessing the gas exchange fitting. The apparatus according to claim 15, wherein the secondary container or the secondary lid further comprises a moisture exchange accessory. P525 18. The apparatus according to claim 7, further comprising: (a) a secondary container capable of enclosing the media storage container when it is attached to the lid; and (b) a secondary lid that attaches to the secondary container. The apparatus according to claim 18, wherein the secondary container or the secondary lid further comprises an access accessory to the means for accessing the media exchange accessory. The apparatus according to claim 18, wherein the secondary container or the secondary lid further comprises a gas exchange fitting. The apparatus according to any of claims 13-20, further comprising a base attachment within the base of the secondary container that is joined with a key mechanism at the base of the media storage container. 22. A method for storing a BAO comprising the steps of: (a) charging the BAO in the apparatus in any of claims 1-8 or 13-20. (b) add a volume of fluid media to the P52.r? media storage container such that the BAO is immersed within the container and the container is substantially filled. 23. A storage method according to. Claim 22, further comprising the step of periodically filling the fluid media. 24. The storage method according to claim 22 further comprises the step of periodically introducing oxygen into the fluid media. 25. A method for transporting a BAO, comprising the steps of (a) charging the BAO in the apparatus of any of claims 1-8 or 13-20. (b) adding a volume of fluid media to the media storage container such that the BAO is immersed within the container and the container is substantially filled; and (c) ship or ship the device to the deployment destination.