WO1999037233A1

WO1999037233A1 - Method and apparatus for aseptically packaging and dispensing live material

Info

Publication number: WO1999037233A1
Application number: PCT/US1999/001316
Authority: WO
Inventors: Daniel Omstead; Barry Weitzner; Antonio Melo; Desmond White
Original assignee: Reprogenesis, Inc.
Priority date: 1998-01-23
Filing date: 1999-01-22
Publication date: 1999-07-29
Also published as: AU2464399A; JP2002500917A; CA2318624A1; EP1049415A1; AU776082B2

Abstract

A method and apparatus for aseptically packaging and dispensing live material are disclosed. The apparatus provides a duplex package comprising an inner container and an outer container completely enclosing the inner container. An access portal transits the outer container and is operative to aseptically introduce a labile fluid or solid from outside the outer container. The inner container is detachably connected to the access portal. Additionally, the outer container is removable. After sterilizing the duplex package, a labile fluid or solid can be introduced through the access portal into the inner container. The access portal can be sealed and the duplex package can be introduced into a sterile environment. The outer container can be removed in the sterile environment. Then a sterile operator can remove the inner container and dispense the labile fluid or solid.

Description

METHOD AND APPARATUS FOR ASEPTICALLY PACKAGING AND DISPENSING LIVE MATERIAL

BACKGROUND OF THE INVENTION 1. Field of the Invention

This invention relates generally to the field of storing and using living material therapeutically. More particularly, the invention relates to a method and apparatus for aseptically packaging and dispensing live material. 2. Description of Related Art In surgical procedures it is routinely necessary to provide sterile surgical instruments. Apparatuses and methods of sterilely introducing non-living material into a sterile field are known in the art, such as disclosed in United States Patent No. 4,065,816, to Sawyer, herein incorporated by reference. Sawyer discloses a method of packaging a prosthetic device for surgical use. The prosthetic device is packaged, sterilized in the package, and shipped to a sterile environment where the package is opened.

While these prior apparatus and methods have been successful with non-living material, they have not succeeded in translocating living material from off-site to a sterile field. Non-living material can be sterilized after being placed into a container, but living materials are adversely affected by the sterilization process and thus have to be treated differently.

Living material can be introduced in a sterile environment, although it is difficult. An example of sterile surgical use of living material is blood transfusion bags. Apparatuses and methods of sterilely using blood bag systems are known in the art, such as disclosed in United States Patent No. 3,942,529, to Waage, herein incorporated by reference. Waage discloses a package and method for storing blood. While blood bags are sterile before they are filled, after filled they are no longer sterile and extreme precaution has to be used once the bags are brought into a sterile environment. Aseptic transfer is an alternative method for introducing living material into a sterile operating room. In this method, the living material is aseptically packed in a vial. The outside of the vial is typically clean, but is not sterile. Therefore, to use the living material in the vial in the operating room, a non- sterile assistant has to open the vial and hold it for the sterile operator. The sterile operator would then draw the cells via a straw or other tube into a sterile syringe. Only then can the living material be used in the sterile environment.

The current method of translocating living material into a sterile operating room is cumbersome and requires additional time and manpower to use the living material in a sterile manner. Therefore, a need has arisen for a new method and apparatus for aseptically packaging and dispensing live material that overcome the disadvantages and deficiencies of the prior art. SUMMARY OF THE INVENTION

In accordance with a principle aspect of the present invention, a method and apparatus for sterilely storing and dispensing live material is disclosed. It is an object of this invention to allow living material to be aseptically introduced into a sterile field.

It is another object of this invention to provide an apparatus capable of sterilely storing and dispensing living material.

These and other objects are met by one or more of the following embodiments. In one embodiment, the invention provides a duplex package comprising an inner container and an outer container completely enclosing the inner container. An access portal transits the outer container and is operative to introduce a labile fluid or solid from outside the outer container into the inner container without further breaching the integrity of the outer container. The access portal allows the labile fluid or solid to be put in the inner container without otherwise opening the outer container. The inner container is preferably detachably connected to the access portal. Additionally, the outer container is removable.

In another embodiment, the invention discloses a method for providing a fluid to a sterile field. The method involves sterilizing a duplex package, the duplex package comprising an outer container that completely encloses an inner container and a fluid-conducting conduit that transits the outer container, with the outer container being removable. The method further involves inserting a fluid from outside the outer container into the sterilized inner container through the fluid-conducting conduit such that the fluid does not breach the sterility of the outer container. The fluid typically comprises material which is labile to sterilization. such as desired living cells suspended therein and the fluid is usually substantially free of contaminant cells other than the desired living cells. This method provides for opening the outer container in proximity to a sterile field, and dispensing the fluid from the inner container. In accordance with this method, once the fluid is inserted into the duplex package, the package is not subject to further gross sterilization.

In another embodiment, the invention discloses a method for storing a labile fluid or solid. The method involves sterilizing a duplex package comprising an outer container that completely encloses an inner container where a sealable access portal transits the outer container. The outer container is removably sealed about the inner container. The method subsequently involves inserting a labile fluid or solid from outside the outer container into the inner container through the access portal, such that the labile fluid or solid does not breach the sterility of the outer container. The package and the labile fluid or solid are not subject to terminal sterilization subsequent to inserting the labile fluid or solid. In another embodiment, the invention discloses a method for sterilely dispensing a labile fluid or solid. The method involves providing a duplex package that comprises an outer container completely enclosing an inner container having a labile fluid or solid therein, with the inner container being detachably connected to an access portal that transits the outer container. The outer container is removable. The method also involves opening the outer container in proximity to a sterile environment and then dispensing the labile fluid or solid from the inner container.

Use of the duplex package of this invention permits a sterile operator to easily handle and use living material in a sterile environment while not risking contamination of the material, the sterile environment, or the sterile operator. In particular, the duplex package of this invention is well suited for containing and presenting implants and cell-containing compositions used in tissue engineering.

Other objects and advantages of the invention are set forth in part in the description which follows, and in part, will be apparent from this description, or may be learned from the practice of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

For a more complete understanding of the present invention, the objects and advantages thereof, reference is now made to the following descriptions taken in connection with the accompanying drawings in which: Figure 1 A is a top perspective of a duplex package according to one embodiment of the present invention.

Figure IB is a side perspective of a duplex package according to one embodiment of the present invention.

Figure 1 C is an end perspective of a duplex package according to one embodiment of the present invention.

Figure 2 illustrates a duplex package with multiple inner containers according to another embodiment of the present invention.

Figure 3 illustrates a duplex package with a thermoformed rigid tray outer container according to another embodiment of the present invention. Figure 4 illustrates a duplex package with a flexible package outside an inner pouch according to another embodiment of the present invention.

Figure 5 illustrates a duplex package with multiple outer containers according to another embodiment of the present invention.

Figure 6 is a perspective representation of a duplex package according to another embodiment of the present invention.

Figure 7 illustrates an example of the duplex package of Figure 6 in use.

Figure 8 illustrates another example of the duplex package of Figure 6 in use. Figure 9 is perspective representation of a duplex package according to another embodiment of the present invention.

Figure 10 is perspective representation of a duplex package according to another embodiment of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

Typically, sterility is ensured for materials coming into a sterile environment, such as an operating room, by subjecting the materials to harsh conditions that kill any adventitious organisms. Such conditions include sterilizing heat, ionizing radiation, toxic gas, etc. When the desired materials include living cells, or easily denatured proteins, nucleic acids, or other organic molecules, alternative procedures for handling these labile materials are needed.

This invention encompasses a method and apparatus for providing a living material to a sterile environment in a faster, easier, and improved manner. The apparatus includes a sterile inner container that is enclosed by an outer container which can be handled by a non-sterile operator or assistant without comprising the sterility of the inner container. To use the apparatus, the inner container is aseptically filled with a living material in an aseptic manner. The apparatus is transported to a non-sterile environment and handled by the non-sterile assistant. The non-sterile assistant can place the apparatus in proximity to a sterile field and remove the outer container and a sterile operator can handle the sterile inner container without fear of breaching the sterility of the material, the operator, the field, or the inner container.

The present invention is particularly beneficial for handling a labile fluid or solid, i.e., a material that undergoes detrimental chemical, physical, or biological changes or breakdown under sterilizing conditions. The labile fluid or solid of the various embodiments disclosed herein may be many things. For example, it may be cells, a cell-containing composition, blood, lymph, or an aqueous suspension containing a heat-denaturable material such as a therapeutic protein in an aqueous solution. The labile material may also be a solid, semi-solid, liquid, sheet (thin or thick), block, other three-dimensional structure, gel, or hydrogel such as particularly or completely hardened alginate. Preferably, the labile material will have cells distributed on the surface and/or throughout all or a portion of the material. The labile fluid or solid may also be cells by themselves, such as a block or sheet of living tissue. Typically, the labile fluid or solid will be a cell-containing composition. In a preferred mode, the duplex package of this invention is used to contain cell-containing compositions for implantation or tissue engineering as described in European Patent Publication Nos. 0299010 and 0469070; International Patent Publication No. WO 94/25080; or United States Patent No. 5,667,778, all herein incorporated by reference. Additionally, a material that could be used, especially in the embodiment depicted in Figure 10, could be the composition depicted in United States Patent No. 5,567,612, herein incorporated by reference. Alternatively, the inner container may contain a fluid composed of living or nonliving material. Typically, such material will be labile to the sterilization method used to sterilize the duplex package. Further, the duplex package disclosed in these embodiments can be used whether sterilized or non-sterilized.

To promote a more complete understanding of the embodiments of the present invention and its advantages, embodiments are discussed in more detail by referring to Figures 1 through 10 of the drawings, like numerals being used for like and corresponding parts of the various drawings. Apparatus of the Present Invention

Referring particularly to Figure 1 A, there is shown a duplex package according a first embodiment of the present invention. The duplex package, indicated generally at 10, comprises an inner container 20 which is completely enclosed by an outer container 22. Outer container 22 has an end that contains a peelable element 32. The other three sides of outer container 22 are sealed at sealed edges 31. A fluid-conducting conduit 24 transits outer container 22. By transiting outer container 22, fluid-conducting conduit 24 allows something outside of outer container 22 to enter inside of outer container 22 without opening outer container 22. Fluid-conducting conduit 24 has an end 30 which is typically closed by a removable closure. For example, the closure at end 30 can include a luer connector, an opening, a cap, or a septum.

Fluid-conducting conduit 24 is detachably connected to inner container 20 at connection point 26. Because inner container 20 is detachably connected, inner container 20 can easily be removed from fluid-conducting conduit 24. The detachable connection may be, for example, a friction fit or luer connector. A labile fluid or cell-containing composition may be inserted into fluid-conducting conduit 24 at end 30 of fluid-conducting conduit 24 such that the labile fluid or cell- containing composition does not result in a breach of outer container 22 that would compromise sterility; the labile fluid or cell-containing composition originating from outside of outer container 22 ends up substantially inside of inner container 20, with little or preferably none of the cell-containing composition of fluid between the outside of inner container 20 and the inside of outer container 22. Thus, introduction of the labile fluid or cell-containing composition does not cause a breach of sterility inside of outer container 22. Further, the labile fluid or cell-containing composition is introduced into inner container 20 without otherwise opening outer container 22.

As shown in Figure IB, a side view of duplex package 10 is depicted. Outer container 22 is formed of a lower layer 34, an upper layer 36, the upper and lower layers 34 and 36 being sealed at three sealed edges, and peelable element 32. Peelable element 32 can be removed to expose inner container 20. Peelable-element 32 can be formed of any material that facilitates easy removal. For example, peelable element 32 can be made of Tyvek® (100% high density polyethylene sheet formed from flash-spun plexifilaments), paper, nylon, or foil. Peelable element 32 can also be a breathable or nonbreathable material. Nonbreathable peelable material is usually made from a plastic. Nonbreathable peelable element is typically two layers coated with an adhesive suitable for separation of the two layers of the peelable element to facilitate removal. Breathable peelable material can either be coated or non-coated. A non-coated peelable material typically generates a stronger seal, but is harder to open. Alternatively, peelable element 32 may be integrally 8 connected to one or more of the upper and lower layers 34 and 36 such that opening peelable element 32 leads to peeling apart the upper and lower layers 34 and 36 at peelable sealed edges 31, shown in Figure 1 A. Further, peelable element 32 may be a tear strip, as described in United States Patent No. 4,367,816, to Wilkes, herein incorporated by reference. Moreover, peelable element 32 may be a sealed edge that can be easily ripped or cut.

Referring again to Figure IB, fluid-conducting conduit 24 provides access to inner container 20 without breaching the seal of outer container 22 at 38. Preferably, the seal at 38 forms a strong seal around fluid-conducting conduit 24, to prevent leakage, such that access to inner container 20 is blocked from the outside of outer container 22, except for fluid-conducting conduit 24.

Fluid-conducting conduit 24 may be, for example, a septum port or a tube. In a typical embodiment, fluid-conducting conduit 24 is made from polyethylene. However, fluid-conducting conduit 24 can be made of any material that can be sealed to outer container 22. For example, such materials are, but not limited to, polyethylene terephthalate glycol, polyvinyl chloride, polystyrene, polyethylene terephthalate, polyethylene, polyester (such as Mylar®), Sclair® (linear low density polyethylene), Tyvek®, paper, nylon, or foil. In an alternative embodiment, fluid-conducting conduit 24 may be substituted by an access portal to facilitate the introduction of a labile fluid or solid comprising larger particles into inner container 20.

In Figure 1C, an end view of duplex package 10 is depicted. Lower layer 34 and upper layer 36 are shown to connect at sealing element 32. End 30 of fluid-conducting conduit 24 is depicted, where a labile fluid or cell-containing composition would be introduced. A luer fitting could be fitted onto end 30 to facilitate easy connection to a syringe or some other device for introducing the labile fluid or cell-containing composition. Alternatively, end 30 could be a septum for inserting a needle into fluid-conducting conduit 24 or an access conduit that can allow the labile fluid or cell-containing composition to be introduced aseptically. End 30 of fluid-conducting conduit 24 is ordinarily sealed to prevent leakage and maintain the sterility of inner container 20. Means for sealing end 30 of fluid-conducting conduit 24 can include any method of sealing end 30 such that a barrier exists between fluid-conducting conduit 24 and the environment of outside outer container 22. For example, sealing means may include a heat seal of end 30 to completely seal end 30, a clasp that clamps shut end 30, a cap for end 30, a fluid seal that seals end 30, or a valve to seal end 30.

In the embodiment depicted in Figures 1A to 1 C, inner container 20 is a syringe; however, inner container 20 can be any containing device. Inner container 20 may be made of any suitable rigid material, such as metal, glass, polypropylene, polycarbonate, or polyethylene. Preferably, inner container 20 will be constructed to facilitate dispensing the labile fluid or cell-containing composition on demand. For example, inner container 20 can be a bag made of any flexible material, and the contents dispensed by squeezing the bag. Any of the components of duplex package 10 can be substituted for any of the like components described with respect to the other embodiments disclosed herein.

In Figure 2, a duplex package 110 containing multiple inner containers is depicted. In addition to inner container 120, a second inner container 140 is also completely enclosed by outer container 122. Inner container 120 has an end 142 and inner container 140 has an end 144. Ends 142 and 144 can be alternatively connected to end 126 of fluid-conducting conduit 124. Additional inner containers can be introduced into outer container 122. Further, additional elements can be enclosed within outer container 122, such as needles, canulae, dispensing valves, bottles, wash fluid, fittings such as a stopcock, connectors, or delivery catheters and mechanisms. Additionally, any other elements that would assist in the delivery of the labile fluid or cell-containing composition, or could easily be assembled and fit within outer container 122, could also be enclosed within outer container 122. In particular, elements that are to be used in the sterile field in conjunction with administration of the labile fluid or cell-containing composition 10 may be enclosed within outer container 122, where they are maintained in sterile condition and readily available at the time the labile fluid or cell -containing composition is dispensed.

A tray 146 can be used as a frame inside of outer container 122 to hold inner containers 120 and 140. Tray 146 is molded to the shape of inner container 120 and 140 to hold inner containers 120 and 140. Use of tray 146 not only keeps inner containers 120 and 140 from being jostled during transit, but also provides an easy method of removing inner containers 120 and 140. For example, tray 146 may be bendably distortable so that when outer container 122 is removed, tray 146 can be contorted to force out inner containers 120 and 140 successively or simultaneously without directly touching inner containers 120 and 140. While tray 146, as depicted, can maintain one or two inner containers, the structure of tray 146 can be altered to maintain any number of inner containers. Tray 146 can be made of any rigid material, for example polyethylene terephthalate glycol, polyvinyl chloride, polystyrene, polyethylene terephthalate, or polyethylene, and preferably tray 146 is elastically distortable. Additionally, all of the components of duplex package 110 can be substituted for any of the like components described with respect to the other embodiments.

In Figure 3, a duplex package 210 is depicted including an outer container 250 comprising a thermoformed tray 252 and a peelable element 232. Thermoformed tray 252 provides a sturdy outer container to protect inner container 220 and maintain the sterility of inner container 220. Peelable element 232 can be easily removed to expose inner container 220. To remove peelable element 232, a user needs only to grasp thermoformed tray 252 and pull outward on peelable element 232. This will separate peelable element 232 from thermoformed tray 252, exposing inner container 220. Additionally, thermoformed tray 252 can be shaped to hold inner container 220, with a tray analogous to tray 146. Thermoformed tray 252 can be made of any sturdy material, including but not limited to polyethylene terephthalate glycol, polyvinyl chloride, polystyrene, polyethylene terephthalate, or 1 1 polyethylene. Peelable element 232 can be formed of any material that facilitates easy removal. For example, peelable element 232 may be made of Tyvek®, paper, nylon, or foil. Alternatively, thermoformed tray 252 may be substituted by a sheet of flexible material, mentioned herein, sealed to peelable element 232 to form outer container 250. Variations on these structures using combinations of flexible and rigid material will be readily apparent to those skilled in the art, and are within the contemplation of this invention (see, e.g.. Figure 4).

A hermetic seal 238 forms a strong seal around fluid-conducting conduit 224. End 230 allows entry of material from outside outer container 222 to inside of inner container 220 via fluid-conducting conduit 224. For example, end 230 can be a check valve or stopcock. Additionally, each of the components of duplex package 210 can be substituted for any of the like components described with respect to the other embodiments.

As shown in Figure 4, a duplex package 310 includes an optional flexible package 362 surrounding an inner pouch 364. Inner pouch 364 is analogous to outer containers 22, 122, and 222 of Figures 1 to 3 and surrounds inner container 320. Inner pouch 364 has a built-in rigid member 366 supporting a fluid-conducting conduit 324. Flexible package 362 provides a flexible container that protects inner pouch 364 and inner container 320 and helps maintains the sterility of inner pouch 364 and inner container 320. Typically, duplex package 310 is sterilized. Then, in a sterile environment, flexible package 362 can be removed and inner container 320 can be filled with a labile fluid or cell-containing composition. This leaves the sterile inner container 320 filled with the labile fluid or cell-containing composition surrounded by inner pouch 364. Inner pouch 364 covers inner container 320 and holds rigid member

366. Flexible package 362 and inner pouch 364 can be made of any flexible material, for example, polyethylene, polyester (such as Mylar®), Sclair®, Tyvek®, paper, nylon, or foil. Inner pouch 364 and flexible package 362 can each include a peelable element to facilitate easy opening. Rigid member 366 can be made of any 12 rigid material mentioned herein. Rigid member 366 assists in forming a strong seal around fluid-conducting conduit 324. End 330 allows entry of material from outside inner pouch 364 via fluid-conducting conduit 324. End 330 can be a can be covered with an end cap 390 or other capping or sealing mechanism. As shown, inner container 320 has an optional connector 396 that couples septum 392 to inner container 320. A user can manipulate inner container 320, with septum 392 at its end, from outside of inner pouch 364 to fit into needle housing 398 so that needle 394 penetrates septum 392. After removing flexible package 362, a labile fluid or cell-containing composition may be inserted through fluid-conducting conduit 324 into inner container 320. After filling inner container 320, septum 392 can be removed from needle 394, separating inner container 320 from fluid conducting conduit 324. Inner container 320 could also be entirely disassembled from connectors, such that a user could assemble inner container 320 and connectors as needed. Additionally, multiple connectors and fittings, preferably having male/female luer connectors, could be used to facilitate connection between inner container 320 and fluid-conducting conduit 324. Moreover, each of the components of duplex package 310 can be substituted for any of the like components described with respect to the other embodiments.

Further, duplex package 310 could be surrounded by a rigid exterior tray, such as tray 146 depicted in Figure 2, such that after removing flexible package 362 and after filling inner container 320, inner pouch 364 enclosing inner container 320 could be held into place by the rigid tray. Alternatively, duplex package 310, before flexible package 362 has been removed, could be inserted into a rigid exterior tray. Using a rigid exterior tray could assist in the shipping, mechanical manipulation, or protecting of inner pouch 364 and inner container 320.

In Figure 5, a duplex package 410 with multiple outer containers is depicted. A first outer container 470 is similar to outer container 250. First outer container 470 includes a thermoformed tray 472 and a peelable element 474, that are similar to thermoformed tray 252 and peelable element 232, respectively. A second 13 outer container 480 completely encloses first outer container 470. Second outer container 480 includes a thermoformed tray 482 and a peelable element 484, that are similar to thermoformed tray 252 and peelable element 232, respectively. However, fluid-conducting conduit 424 extends through the multiple outer containers to the outside of second outer container 480, allowing a labile fluid or cell-containing composition to be introduced from outside of second outer container 480 into inner container 420. Having multiple outer containers provides additional protection and sterility. Duplex package 410 can have any number of outer containers as long as fluid-conducting conduit 424 extends through the outermost outer container. In fact, any of the components of duplex package 410 can be substituted for any of the like components described with respect to the other embodiments.

In Figure 6 duplex package 510 according to another embodiment of the present invention is depicted. Outer container 522 completely encloses inner container 520, and tray 546 holds inner container 520 in place. While duplex package 510 is similar to duplex package 10, a significant difference between the two embodiments is that outer container 522 has opening 556 that provides controlled access to the interior of outer container 522. Typically opening 556 will be sealed before and after filling of inner container 520 with a labile fluid or cell- containing composition. End 542 of inner container 520 can be lined up with opening 556 for the filling operation. End 542 can include an opening, a septum, a luer connector, or a stopper. For example, end 542 can be made of cork, rubber, or plastic. As depicted, fluid-conducting conduit 524 is a hollow needle connected to a syringe 590, which is part of the filling apparatus. Syringe 590 can be filled with a suitable fluid which may be a cell-containing composition or may contain other material that is labile to sterilization. Fluid-conducting conduit 524 is detachably connected to inner container 520 during the filling operation, for needle 524 passes through or into end 542 of inner container 520 and is removable after filling inner container 520. Additionally, any of the components of duplex package 510 can be 14 substituted for any of the like components described with respect to the other embodiments.

After inserting the labile fluid or cell-containing composition into inner container 520, needle 524 can be removed. To seal outer container 522, opening 556 can be sealed with sealing device 558, as shown in Figure 7. For example, sealing device 558 can be a cap, a luer connector, a valve, a fluid valve, a clamp that shuts opening 556, or a heat seal such that opening 556 is heat sealed shut.

To maintain sterility of duplex package 510 after sterilization and before filling inner container 520, an outer sterile barrier 564 can be enclosed entirely around outer container 522 as shown in Figure 8. Outer sterile barrier 564 can be, for example, made of any material that can maintain the sterility of outer container 522 and inner container 520. For example, polyethylene, low density polyethylene, polyester (such as Mylar®), Sclair®, Tyvek®, paper, nylon, or foil can be used. Additionally, outer sterile barrier 564 can be rigid or flexible, thus, for example, polyethylene terephthalate glycol, polyvinyl chloride, polystyrene, or polyethylene terephthalate, could also be used. Typically, package 510 would be sterilized with barrier 564 in place, and barrier 564 would be removed immediately prior to filling inner container 520 with material that is labile to sterilization. Operation of the Present Invention

In operation, the duplex package of all the embodiments function similarly. The duplex package is sterilized by a known sterilization means. Typically, the duplex package is filled aseptically using a sterile filling apparatus, preferably involving a biosafety cabinet. To fill the inner container, a labile fluid or solid is inserted from outside of the outer container into the inner container through an access portal, which may be a fluid-conducting conduit. The access portal allows the labile fluid or solid to be introduced into the inner container without otherwise opening the outer container. Additionally, the labile fluid or solid does not breach the sterility inside of the outer container. Thus, the labile fluid or solid does not substantially enter the area between the outside of the inner container and the inside 15 of the outer container. The labile fluid or solid may or may not be sterile. Where the labile solid or fluid contains a desired population of living cells, but otherwise is substantially free of contaminant cells other than those living cells, the only non- sterile material in the sterile duplex package would be those desired living cells. If, however, the labile solid or fluid does not contain living cells, but rather non-living labile material, then everything in the duplex package may be sterile.

After inserting the desired labile fluid or solid into the inner container, any excess labile fluid or solid left in the access portal or fluid-conducting conduit can be pushed into the inner container by using some means to remove the excess labile fluid or solid. Any means for purging the access portal or fluid- conducting conduit of excess labile fluid or solid is acceptable. For example, the excess material can be pushed using a rod, by forcefully blowing air into the access portal or fluid-conducting conduit, or by tapping the access portal or fluid- conducting conduit. The end of the access portal or fluid-conducting conduit may then be closed. Any sealing means may be used as long as a good seal is created that keeps the inner container sterile. The duplex package may then be translocated to a sterile field, such as an operating room.

The duplex package, once filled with the labile fluid or solid, is not further subjected to terminal sterilization. Terminal sterilization would kill everything living in the duplex package. Because the labile fluid or solid preferably contains living material or other materials labile under sterilizing conditions, terminal sterilization would adversely affect the labile fluid or solid. Therefore, gross sterilization of the package is only performed prior to insertion of the labile fluid or solid. A non-sterile operator or assistant may then take the duplex package and bring it into the operating room or adjacent to a sterile field. The non-sterile assistant can remove the outer container of the duplex package. To remove the outer package, the non-sterile operator might simply peel off the peelable element of the outer container, tear or rip open the outer container, or cut the outer container. Once the outer container has been opened, a sterile operator can grasp the inner container 16

(or one of them if there are several) and disconnect it from the access portal or fluid- conducting conduit, if it is attached. Alternatively, if the inner containers are contained by a tray, the non-sterile assistant can bend the tray such that an inner container pops out into the sterile field or the sterile operator's hand. Alternatively still, the non-sterile assistant can dump the inner containers out of the outer container, once the outer container has been opened. Alternatively still, if the inner container is a pouch, the sterile operator can grasp the pouch. Alternatively still, if using a live material, such as a skin sample, the non-sterile assistant can open the inner container and let the sterile operator grasp the sample. Alternatively still, if using a sample that has solidified in the inner container to a desired shape, the sterile operator could open the inner container and remove the sample. Typically the solidified sample will be a cell-containing composition or material, such as a preformed implant; however, it is within the contemplation of this invention that the preformed implant contain material labile to sterilizing conditions, such as a therapeutic protein, but no cells. The outer container according to this invention can be removed or opened such that inner container is not contaminated and the inner container is not exposed to any non-sterile surface.

Once the sterile operator has the inner container, the sterile operator can dispense the labile fluid or solid. If the inner container is a syringe, the syringe plunger can be pushed to dispense the contents. If the inner container is a pouch, the inner container can be squeezed to release the desired labile fluid or solid.

To more completely describe the method of the present invention, the method will be further described with relation to the particular embodiment depicted in Figures 1A to 1C. However, the method as described below could be readily adapted for operation with the duplex package of any of the embodiments disclosed herein. End 30 of fluid-conducting conduit 24 reversibly is sealed. Means to seal end 30 include, for example, heat sealing the end, a cap, a fluid valve, a value, or clamping shut end 30. Duplex package 10, with end 30 sealed, is then sterilized to kill substantially all contaminating organisms in duplex package 10. Preferably, residual contamination will be at or below the level acceptable under good 17 manufacturing practices for medical devices. Duplex package 10 could be sterilized with any suitable method, for example, by steam, ethylene oxide, gamma irradiation, electron beam irradiation, or with ultraviolet light.

After being subjected to sterilization by a suitable means, inner container 20 is filled aseptically. In one example, duplex package 10 is transported to a biosafety cabinet, such as a class 100 sterilization hood. Once inside of the biosafety cabinet, the removable seal on end 30 of fluid-conducting conduit 24 is removed. The labile fluid or cell-containing composition is then inserted into inner container 20 of duplex package 10. The labile fluid or cell-containing composition may be inserted by any one of numerous methods. In one embodiment, a sterile filling syringe, with a luer lock is connected to end 30 of fluid-conducting conduit 24. This filling syringe can be connected manually or mechanically, such that sterility in the interior of the biosafety cabinet is not breached. The filling syringe is then emptied into fluid-conducting conduit 24 and the labile fluid or cell-containing composition is forced into inner container 20. After filling inner container 20, the filling syringe is removed. Optimally, any excess labile fluid or cell-containing composition left in fluid-conducting conduit 24 can be pushed into inner container 20 by using any means for purging fluid-conducting conduit 24 of excess fluid, for example by using a rod to push the excess fluid into inner container 20, by forcefully blowing air into fluid-conducting conduit 24, or by tapping fluid-conducting conduit 24. After the labile fluid or cell-containing composition has been inserted into inner container 20, end 30 of fluid-conducting conduit 24 is then sealed again with any of the above methods. End 30 can even be permanently sealed with heat. At this point, duplex package 10 is not subsequently subjected to terminal sterilization that would affect the labile fluid or cell-containing composition.

Terminal sterilization is a final sterilization that substantially kills all living things prior to shipment of a sterile product. Accordingly, terminal sterilization would adversely affect the labile fluid or cell-containing composition after inner container 20 of duplex package 10 has been filled. However, so long as 18 the labile fluid or cell-containing composition is substantially free of contaminants other than the desired cells, and inner container 20 is sterile prior to inserting the labile fluid or cell-containing composition, once the labile fluid or cell-containing composition is inserted aseptically, inner container 20 will be substantially free of living cells except for the desired cells that are completely enclosed in inner container 20. Of course, if the fluid only contained nonliving material, duplex package 10 could be subject to terminal sterilization after the nonliving material is inserted into inner container 20, unless the non-living material was labile under the conditions of the terminal sterilization. Duplex package 10 is then typically transported to the point of use, which may be an operating room with a sterile environment. At this point, the inside of outer container 22 and the outside of inner container 20 will be sterile (i.e., substantially free of contaminating organisms). Additionally, the inside of inner container 20 will be sterile, except for the desired labile fluid or cell-containing composition. A non-sterile assistant in the operation room can peel open outer container 22, exposing inner container 20 without contaminating it. A sterile operator can then grasp inner container 20 and twist inner container 20 free from the luer connector. Alternatively, if tray 46 was used, the non-sterile assistant could remove outer container 22 and push inner container 20 free from the tray 46 by indirect pressure without contaminating the outside of inner container 20, allowing inner container 20 to fall either into the sterile environment or the sterile operator's hand. Alternatively, if inner container 20 was no longer attached to fluid-conducting conduit 24, outer container 22 could be removed by the non-sterile assistant without touching inner container 20 directly, and the sterile operator could either grasp inner container 20 or let it fall out of outer container 22 into the sterile field.

After removing inner container 20, the sterile operator can use inner container 20 to dispense the labile fluid or cell -containing composition. For example, if inner container 20 is a syringe, the labile fluid or cell-containing composition can be dispensed through the syringe. Alternatively, if inner container 19

20 is a flexible bag, inner container 20 can depressed to squeeze or squirt out the labile fluid or cell-containing composition.

It will be noted that everything contained within outer container 22 and translocated to the sterile environment will typically be in sterilized and in clean condition, previously untouched until grasped by the sterile operator. Further, if inner container 20 is a syringe or a bag, then only the exterior of inner container 20 would be touched by the sterile operator. The needle, cell -containing composition, or fluid would typically be entirely untouched by any operator. Thus, the labile fluid or cell-containing composition is aseptically introduced to the sterile field or patient. Methods for using other embodiments of this invention for manipulation of labile fluids or solids in sterile environments will be analogous to the method described in detail for use of the embodiment of Figure 1 with a cell- containing composition. Alternative Embodiments of the Present Invention As shown in Figure 9, a duplex package 610 according to another embodiment of the present invention is depicted. An outer container 622 completely encloses an inner container 620, and a tray 646 holds inner container 620 in place. While duplex package 610 is similar to duplex package 510, the significant difference between the two embodiments is that outer container 622 has a flap 664. Flap 664 includes an access portal 624. Access portal 624 can be aligned with an end 642 of inner container 620. Access portal 624 allows a labile fluid or solid, to enter from outside of outer container 622 and into inner container 620. Additionally, any or all of the components of duplex package 610 can be substituted for any of the like components described with respect to the other embodiments. In operation, duplex package 610 may operate as follows. End 642 of inner container 620 is not originally connected to access portal 624, but rather inner container 620 is free floating within outer container 622. Duplex package 610 is sterilized and then translocated to a filling station, for example a biosafety cabinet. While in the biosafety cabinet, end 642 can be connected to access portal 624. End 20

642 can be connected using any known connecting means. Preferably, outer container 622 is a flexible material which may be manipulated to align end 642 with access portal 624. After sterilizing duplex package 610 and connecting end 642 to access portal 624, a labile fluid or solid is inserted from outside of outer container 622 to inside of inner container 620. After inserting the labile fluid or solid, end 642 is disconnected from access portal 624 and access portal 624 is closed. Access portal 624 can be sealed by heating outer container at the area marked 668, or fusing closed access portal 624. Alternatively, it can be sealed with any method that provides a barrier to contamination of the sterility of inside outer container 622, for example, with a clamp, by melting the ends of access portal 624 in some other manner, with a cap, a valve, a fluid valve, or stopper. Accordingly, inner container 620 is free floating within the completely sealed outer container 622. Alternatively, the tip of end 642 can be fused or heated shut along with access portal 624. In that case, inner container 620 would be connected to the completely sealed outer container 622. Regardless, access portal 624 is removed and is not longer part of duplex package 610.

Duplex package 610 is then translocated to a location adjacent to a sterile environment or field. Once in or near the sterile environment or field, a non- sterile assistant can remove outer container 622. One such method could be to cut, tear, or rip along dotted line 676, removing flap portion 668. However, outer container 622 can be removed in any conventional manner as long as the non-sterile assistant does not come into substantial contact with inner container 620. After removing outer container 622, a sterile operator can grasp inner container 620, if inner container 620 has a graspable portion that is easily grasped, or can allow inner container 620 to fall out from outer container 622. If inner container 620 is still connected to outer container 622, inner container 620 can be sterilely removed by any suitable means. The labile fluid or solid can then be sterilely dispensed.

As shown in Figure 10, a duplex package 710 according to another embodiment of the present invention is depicted. Duplex package 710 is well suited 21 to handle solid materials, including tissues such as a living skin sample, but would also function well with any other labile fluid or cell-containing material, for example in the form of sheets (thin or thick), blocks, or other three dimensional structures. Typically the solid material will be a cell-containing material, such as a preformed tissue implant; however, it is within the contemplation of this invention that the solid be a matrix containing material labile to sterilizing conditions, such as a therapeutic protein, but no cells. Inner container 720 may resemble a container for storage and distribution of a skin wound dressing container disclosed in Tubo et al.. U.S. Patent No. 5,040,677, incorporated herein by reference, except that access portal 724 extends into inner container 720. In a typical example, inner container 720 includes thermoformed tray 772 and an peelable element 774. Outer container 722 completely encloses inner container 720, and access portal 724 also transits outer container 722. Alternatively, access portal 724 can be a joining point of inner container 720 and outer container 722, as depicted in Figure 10. Outer container 722 can alternatively be made of a flexible material, as previously disclosed.

Additionally, any or all of the components of duplex package 710, can be substituted for any of the like components described with respect to the other embodiments.

After sterilizing duplex package 710, a sample 790 could be sterilely inserted through access portal 724 in a sterile hood or other environment suited for aseptic filing, and into inner container 720. After inserting sample 790, access portal 724 may be sealed by any sealable means, including the previously mentioned means. After sealing, duplex package 710 may be translocated to a sterile field. Once package 710 is in proximity to the sterile field, a non-sterile assistant can remove outer container 722 by any of the fore-mentioned means, including separating peelable element 784. Then a sterile operator can remove sterile peelable element 774 and remove sample 790.

Alternatively, inner container 720 could be a molded shape. A cell- containing composition or other labile fluid could be inserted into inner container 720 such that once inside, the cell-containing composition or other labile fluid 22 solidifies into the shape of the molded inner container 720. To use, outer container 722 and inner container 720 could be removed, exposing the molded sample to manipulation by the sterile operator.

Alternatively, a seedable substrate could be placed in inner container 720, either before or after the duplex package 710 has been sterilized. If the substrate is placed in inner container 720 before the duplex package 710 is sterilized, the duplex package 710 should be sterilized before living cells are introduced to the inner container 720. Cells or some other cell-containing composition may then be sterilely introduced to the inner container, as described above, to seed the substrate with cells or other cell-containing composition. When the cells, that have now been seeded by the substrate, are to be removed, they may be removed as described above.

While use of the duplex aseptic package of this invention has been described in terms of an operating room containing a sterile field, where the package is opened and used by a non-sterile assistant and sterile operator, respectively, those skilled in the art will recognize that the apparatus and methods for its use are easily adapted to other environments with similar requirements, including a sterile manufacturing facility or a sterile field in an outpatient clinic. The non-sterile assistant may be substituted by mechanical means of opening the outer container, or even by one hand of the sterile operator while preventing contamination of the other hand which remains in the sterile field.

While the invention has been particularly shown and described by the foregoing detailed description, it will be understood by those skilled in the art that various other changes in form and detail may be made without departing from the spirit and scope of the invention.

Claims

23CLAIMS:We claim:

1. A duplex package comprising: an inner container; an outer container completely enclosing said inner container; an access portal transiting said outer container, wherein said access portal is operative to introduce a labile fluid or solid from outside said outer container into said inner container without otherwise opening said outer container; said inner container being detachably connected to said access portal; and means for removing said outer container.

2. The duplex package of claim 1 , wherein said labile fluid or solid is in said inner container.

3. The duplex package of claim 2, wherein said outer container includes an inner side and an outer side, wherein said inner side of said outer container is sterile, further wherein said inner container is substantially free of living cells other than cells contained in said labile fluid or solid.

4. The duplex package of claim 3, wherein said outer container includes a graspable portion such that an operator can hold onto said outer container and remove said outer container without contaminating said inner container.

5. The duplex package of claim 3, wherein operation of said means for removing said outer container does not contaminate said inner container and does not expose said inner container to a non-sterile surface.

6. The duplex package of claim 2, wherein said labile fluid or solid is selected from the group consisting of cells, a cell-containing composition, blood, lymph, an aqueous suspension, a solid, a semi-solid, a liquid, a gel, a hydrogel, and a solid matrix having cells disbursed therein.

7. The duplex package of claim 2, wherein said access portal includes an end, and said labile fluid or solid is retained by heating said end or clasping shut said end. 24

8. The duplex package of claim 2, wherein said access portal includes sealing means that comprises a cap, a fluid seal, or a valve.

9. The duplex package of claim 1 , wherein said access portal includes an end, wherein said end includes a luer fitting.

10. A duplex package comprising: an inner container; an outer container completely enclosing said inner container; a fluid-conducting conduit transiting said outer container, wherein said fluid-conducting conduit is operative to introduce a fluid from outside said outer container into said inner container without otherwise opening said outer container, said inner container being detachably connected to said fluid-conducting conduit; and means for removing said outer container to expose said inner container.

1 1. The duplex package of claim 10, wherein said fluid is in said inner container.

12. The duplex package of claim 1 1, wherein said outer container includes an inner side and an outer side, wherein said inner side of said outer container is sterile, further wherein said inner container is substantially free of living cells other than cells contained in said fluid.

13. The duplex package of claim 12, wherein said outer container includes a graspable portion such that an operator can hold onto said outer container and remove said outer container without contaminating said inner container.

14. The duplex package of claim 12, wherein operation of said means for removing said outer container does not contaminate said inner container and does not expose said inner container to a non-sterile surface.

15. The duplex package of claim 1 1 , wherein said fluid is selected from the group consisting of blood, lymph, an aqueous suspension containing a heat- denaturable material, and a therapeutic protein in an aqueous solution, 25

16. The duplex package of claim 1 1, wherein said fluid forms a solid, a semi-solid, a gel, a hydrogel, or a solid matrix having cells disbursed therein.

17. The duplex package of claim 1 1 , wherein said fluid-conducting conduit includes an end, and said fluid is retained by heating said end or clasping shut said end.

18. The duplex package of claim 1 1 , wherein said fluid-containing conduit includes sealing means that comprises a cap, a fluid seal, or a valve.

19. The duplex package of claim 10, wherein said fluid-conducting conduit includes an end, wherein said end includes a luer fitting.

20. The duplex package of claim 1 or 10, wherein said inner container comprises a syringe.

21. The duplex package of claim 1 or 10, further comprising a plurality of inner containers.

22. The duplex package of claim 1 or 10, wherein at least one of said inner container or said outer container comprises a flexible material.

23. The duplex package of claim 1 or 10, wherein at least one of said inner container or said outer container comprises a rigid material.

24. The duplex package of claim 1 or 10, wherein said means for removing said outer container comprises a peelable element.

25. The duplex package of claim 1 or 10, wherein said inner container is supported by a frame or a rigid tray.

26. The duplex package of claim 1 or 10, wherein said outer container is completely enclosed by an outer sterile barrier.

27. The duplex package of claim 26 further comprising means for removing said outer sterile barrier.

28. Method for providing a fluid to a sterile field comprising: sterilizing a duplex package comprising an outer container that completely encloses an inner container, a fluid-conducting conduit that transits said outer container, said outer container being removable; 26 inserting a fluid from outside said outer container into said inner container through said fluid-conducting conduit such that said fluid does not breach sterility of inside of said outer container, said fluid comprising desired living cells suspended therein, said fluid further being substantially free of contaminant cells other than said desired living cells; opening said outer container in proximity to a sterile field; and dispensing said fluid from said inner container into said sterile field, wherein subsequent to said step of inserting said fluid into said inner container, said duplex package is not subject to terminal sterilization.

29. The method of claim 28, wherein said inner container is detachably connected to said fluid-conducting conduit.

30. The method of claim 28, further including a step of: translocating said inner container to said sterile field independently of said outer container.

31. The method of claim 28, wherein said outer container includes an inner side and an outer side, wherein said inner side of said outer container is sterile, further wherein said inner container is substantially free of living cells other than cells contained in said fluid.

32. The method of claim 28 wherein said duplex package further comprises a plurality of inner containers.

33. Method for storing a labile fluid or solid comprising: sterilizing a duplex package comprising an outer container that completely encloses an inner container, an access portal that transits said outer container, said outer container being removable; and inserting a labile fluid or solid from outside said outer container into said inner container through said access portal such that said labile fluid or solid does not breach sterility of inside of said outer container, wherein said labile fluid or solid is not subject to sterilization subsequent to said step of inserting said labile fluid or solid. 27

34. The method of claim 33 further including a step of: removing excess labile fluid or solid in said access portal.

35. The method of claim 34, wherein said step of removing excess labile fluid or solid uses air, a rod, or tapping.

36. The method of claim 33, wherein said inner container is connected to said access portal by means including a luer fitting.

37. The method of claim 33, wherein said outer container includes a graspable portion, which permits an operator to hold onto said outer container and remove said outer container without contaminating said inner container.

38. The method of claim 33, wherein said inner container is detachably connected to said access portal.

39. Method for sterilely dispensing a labile fluid or solid comprising: providing a duplex package comprising an outer container completely enclosing an inner container having a labile fluid or solid therein, an access portal that transits said outer container, said outer container being removable; opening said outer container in proximity to a sterile environment; and dispensing said labile fluid or solid from said inner container.

40. The method of claim 39, wherein said inner container is detachably connected to said access portal.

41. The method of claim 39, wherein said access portal is a fluid- conducting conduit, and said method further including a step of removing said inner container from said fluid-conducting conduit prior to said step of opening said outer container.

42. The method of claim 39, further including a step of: translocating said inner container to said sterile field independently of said outer container.

43. The method of claim 42, wherein said step of translocating comprises grasping said inner container and removing said inner container. 28

44. The method of claim 43, wherein said step of removing comprises detaching said inner container from said access portal.

45. The method of claim 33 or 39, wherein said outer container includes an inner side and an outer side, wherein said inner side of said outer container is sterile, further wherein said inner container is substantially free of living cells other than cells completely enclosed in said inner container.

46. The method of claim 33 or 39, wherein said labile fluid or solid is selected from the group consisting of cells, a cell-containing composition, blood, lymph, an aqueous suspension containing a heat-denaturable material, a solid, a semi-solid, a liquid, a gel, a hydrogel, and a solid matrix having cells disbursed therein.

47. The method of claim 33 or 39, wherein said access portal includes an end, and said labile fluid or solid is retained by heating said end or clasping shut said end.

48. The method of claim 33 or 39, wherein said access portal includes sealing means that comprises a cap, a septum, a fluid seal, or a valve.

49. The method of claim 33 or 39, wherein said inner container comprises a syringe.

50. The method of claim 33 or 39, wherein said duplex package further comprises a plurality of inner containers.

51. The method of claim 33 or 39, wherein at least one of said inner container or said outer container comprises a flexible material.

52. The method of claim 33 or 39, wherein at least one of said inner container or said outer container comprises a rigid material.

53. The method of claim 33 or 39, wherein said outer container includes a peelable element.

54. The method of claim 33 or 39, wherein said access portal includes an end, wherein said end includes a luer fitting.

55. The method of claim 33 or 39, wherein said inner container is supported by a frame or a rigid tray.