Abstract:
Disclosed is a device useful for preserving tissue samples after extraction from human or animal patients. The device includes a circular blade for taking a tissue sample, the blade being coupled to a sealing cap which mates to one end of a container. A retrieval port is also disclosed for extracting fluids from the container. Methods are also disclosed by which tissue samples may be taken and prepared for storage, processing, culture, or other analysis.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
       [0001]    This application is a Continuation of U.S. Ser. No. 13/571,044 filed on Aug. 9, 2012, which claims the benefit of U.S. Provisional Patent Application Ser. No. 61/521,625 filed Aug. 9, 2011, which is incorporated herein by reference in its entirety. 
     
    
     BACKGROUND 
       [0002]    The present invention relates to a device for extraction, collection, transport, processing and preservation of a living tissue sample. 
         [0003]    Certain therapies utilize stem cells for the treatment of diseases and conditions in humans and animals. Optimally the stem cells are obtained from the patient, cultured and processed for storage. Many stem cell based therapies require a large number of viable cells. Procedures for culturing and preserving a desired population of stem cells have been well refined over the years. However, one common problem that remains is in collecting the cells and transporting them to another location for culturing. 
       SUMMARY 
       [0004]    The embodiments disclosed are directed to tissue extraction and collection devices and methods of using such devices to preserve and retrieve collected samples. The disclosed embodiments include vials having containers and accompanying blades attached to the container caps which are useful together to collect a tissue sample and maintain it for later testing, culturing, processing, or storage. 
         [0005]    Further, still additional embodiments will be apparent to those skilled in the art from the Detailed Description herein. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0006]      FIG. 1  is a view of an overwrap containing the punch biopsy vial of the present disclosure. 
           [0007]      FIG. 2  is an exploded diagram of the vial shown in  FIG. 1 . 
           [0008]      FIG. 3  is a view of the vial shown in  FIG. 2  with a tissue sample contained therein. 
           [0009]      FIG. 4  is a diagram of a step for withdrawing cells from the vial shown in the prior figures. 
           [0010]      FIG. 5  is an enlarged cross-sectional view of the cap and blade for the vial shown in  FIG. 2 . 
           [0011]      FIG. 6  is a cross-sectional view of a modified cap for the vial shown in the prior figures. 
           [0012]      FIG. 7  is another embodiment of the punch biopsy vial shown in  FIGS. 1-4 . 
           [0013]      FIG. 8  is an exploded diagram of the vial shown in  FIG. 7 . 
           [0014]      FIG. 9  is end view of the distal end of the vial shown in  FIG. 7 . 
           [0015]      FIG. 10  is a side view of a handle for use with the biopsy vials shown in  FIG. 1-9 . 
           [0016]      FIG. 11  is an end view of the distal end of the handle shown in  FIG. 10 . 
       
    
    
     DETAILED DESCRIPTION 
       [0017]    For the purpose of promoting an understanding of the principles of the invention, reference will now be made to embodiments, some of which are illustrated in the drawings, and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended. Any alterations and further modifications in the described embodiments, and any further applications of the principles of the invention as described herein are contemplated as would normally occur to one skilled in the art to which the invention relates. 
         [0018]      FIGS. 1-4  show one embodiment of a system useable for tissue extraction procedures in human or animal (veterinary) patients. The tissue extraction system is shown at  10  and it includes a punch biopsy vial  15  that can be provided to any physician in a sterile sealed overwrap  11 , as shown in  FIG. 1 . The overwrap  11  can be, and is preferably, of the type conventionally used for sterile medical devices. The punch biopsy vial  15  includes a container  16  and a cap  20 , as shown in  FIG. 2  and  FIG. 3 . In this embodiment, the container  16  has an open mouth  17  at one end and a retrieval port  18  defined at the other end of the container. In another embodiment shown in  FIGS. 7-9  and described in greater detail below, a retrieval port  77  similar to retrieval port  18  is included within cap  20 . Regardless of position, the retrieval port may have a variety of configurations provided that it can maintain a sterile, sealed, leak-proof closure, while permitting access to withdraw the contents of the container. For instance, the retrieval port  18  may constitute a septum that may be pierced by a syringe, as described herein. Alternatively the port may include a valve arrangement. The retrieval port  18  may be provided with a cover to maintain the sterility of the port until it is desired to access the contents of the container  16 . 
         [0019]    The cap  20  includes a mating interface  21  adapted to sealingly mate with the mouth  17  of the container  16 . The interface may thus be a conventional threaded engagement or Luer-type fitting between the cap and container and may incorporate a seal or gasket between the cap and the container to ensure a fluid-tight seal. The handle  22  of the cap provides a gripping surface to thread the cap onto the container. 
         [0020]    In one aspect, the cap  20  incorporates a circular blade  25  that may be configured similar to blades used for a punch biopsy. In one embodiment, the blade is a sterilized stainless steel blade having a diameter of between about 2 mm and about 4 mm. The blade diameter is preferably calibrated for taking a tissue sample from the skin of the patient. The sample should be sufficiently large to ensure a large quantity of viable stem cells, yet not so large as to cause undue trauma to the patient. It has been found that a sample as small as 2 mm provides a sufficient quantity of viable cells. A sample larger than 4 mm yield more cells but the increase in viable cells is of only minimal advantage when culturing the cell sample. On the other hand, the larger diameter lesion left by a skin punch greater than 4 mm has been found to be uncomfortable and unnecessary. 
         [0021]    The circular blade  25  is embedded within the cap  20  and concentrically disposed inside the mating interface  21 . The blade is solidly anchored within the handle  22  of the cap so that the blade can be rotated by the physician by rotating the cap using the handle. In one embodiment, the blade is mounted within the cap using an over-molding process. The blade may incorporate features to enhance the anchorage of the blade so that it cannot be dislodged from the cap. For instance, as shown in  FIG. 5 , the blade may include a flared end  27  that is embedded within the handle  22 . As further shown in  FIG. 5 , the blade may include serrations or teeth  26  to facilitate severing the tissue, especially upon rotation of the cap  20 . The blade has a length adapted to an appropriate depth for the region of tissue or skin targeted for recover. In one specific embodiment the blade has a height of about 5 mm for an epidermal skin sample. 
         [0022]    In one procedure using the tissue extraction system  10 , the physician removes the vial  15  from the package  11  and releases the cap  20  from the container  16 . The physician then pierces the patient&#39;s skin with the blade  25 , preferably while rotating the blade. When the blade is removed the tissue sample is trapped within the blade. The cap  20  is then remounted onto the vial  16 , as shown in  FIG. 3 . The tissue sample T may be dislodged into a solution  30  contained within the vial, such as by tapping the cap. It is contemplated that the vial may be provided with the solution as part of the sealed overwrap  11 . Alternatively, the solution may be added by the physician, such as from a separate vial provided with the sealed overwrap. However, in order to simplify the procedure it is preferable that the vial be initially filled with the solution  30  so that no problems arise in collecting and transporting the tissue sample T. In one embodiment, the cap  20  may be provided in the overwrap  11  separated from the vial  15 . The vial may contain the solution  30  and be provided with a seal that can be broken or penetrated by the blade  25  when the cap is fastened to the vial. 
         [0023]    In one aspect of the present disclosure, the solution  30  contained within the vial and into which the tissue sample T is placed, may constitute a variety of compositions depending upon the desired function of the sealed vial shown in  FIG. 3 . In one embodiment, the solution may be a standard buffered saline solution, with or without other additives depending upon the nature of the tissue and the amount of time that the tissue sample T will remain in the solution before subsequent processing. The solution may also include a chemical preservative that is capable of preserving the tissue sample but is inert to the cells of interest. Various embodiments of solution  30  include any liquid medium commonly known in the art. 
         [0024]    In another embodiment, the solution  30  may be a preservative adapted to preserve the tissue and its cells intact. The preservative may be a cryoprotectant so that the vial  15 , complete with tissue sample T, may be transported directly to a cryogenic storage facility. The preservative may thus include known permeable cryoprotectant agents, such as dimethyl sulfoxide, ethylene glycol, propylene glycol and glycerol. 
         [0025]    In a further alternative, the solution  30  may incorporate enzymes adapted to digest the tissue sample and liberate cells of interest. The enzymatic activity may be modulated by temperature either immediately once the sample is acquired or later at a separate facility. The enzyme can be selected as a function of the tissue being digested, the desired cell, the digestion rate and/or the optimal time of digestion. In the latter case, the digestion time may be controlled by shipping the sealed vial in a shipping container having a timed cooling feature that is activated at an appropriate time to slow the enzymatic activity. One suitable enzyme is ACCUTASE® sold by Life Technologies Corp dba Invitrogen Corp. 
         [0026]    In this embodiment, it is thus contemplated that the solution will digest the tissue to liberate cells C of interest, as shown in  FIG. 4 . The cells C may be removed from the vial  15  using a hypodermic needle  42  and syringe  40 . The needle penetrates the sealed end  18  of the vial in a known fashion. The suspended cells are withdrawn into the syringe to be taken to other processing. 
         [0027]    In order to facilitate dislodging the tissue sample into the vial, a plunger arrangement may be provided as illustrated in  FIG. 6 . The cutting blade  25  may be the same as described above, mounted within the mating interface  51  of the cap  50 . As shown in  FIG. 6 , the mating interface may be in the form of a tapered stopper, or maybe the threaded interface described above. The cap  50  is longer than the cap  20 , including a longer handle  52 . The cap provides a passageway  54  through which a plunger  60  extends. The plunger  60  includes an end  62  that is depressed to push the plunger down through the blade  25  to dislodge the tissue sample from within the blade. The plunger may include a seal  64 , such as on O-ring, disposed between the plunger and the passageway  54 . In addition, a spring  66  may be provided between the end  62  and the handle  52  to bias the plunger away from the blade. 
         [0028]    Another embodiment of the punch biopsy vial is show in  FIG. 7 - FIG. 9  at  68  which is similar to vial  15 . Biopsy vial  68  includes a container  79  similar to container  16  except that in this embodiment, container  79  has an open mouth  17  at one end and closed end  82  defined at the other end of container  79  rather than a retrieval port. A retrieval port  77  similar in all respects to retrieval port  18  described above is incorporated into a cap  85  which includes the embodiments of handle  22  and mating region  21  described above. Cap  85  is constructed similarly to the embodiments of cap  20  described above and includes a circular blade  25  that is positionable inside container  79  as shown in  FIG. 7 . One embodiment of retrieval port  77  is shown in  FIG. 9  as a septum configured within the central area of cap  85  such that a needle or other similar instrument can be inserted through the septum into container  79  to retrieve the contents of the container similar to what is shown in  FIG. 4  with respect to the previous embodiment. 
         [0029]    Also shown in  FIG. 10  and  FIG. 11  is one embodiment of an elongate handle  70  that may be used in conjunction with the embodiments of the punch biopsy vial described above. The handle has an elongate tubular shape with a contact region  73  at the distal end which in one embodiment can be pressed against the cap of the biopsy vial to aid in steadying blade  25  during a tissue extraction or other procedure. Contact region  73 , in one embodiment, press fits into a corresponding detent, notch, or similar receiving region in the cap to firmly steady handle  70  during the procedure. Handle  70  may also be secured to the cap by screw threads at or near the contact region  73  that correspond with similar threads in the cap. Other nondesctructible attachment means may be used as well such as pins, snaps, and the like. In another embodiment, handle  70  is formed as part of the vial cap and designed to be broken or otherwise separated from the cap and discarded after the tissue extraction procedure has been completed. In still another embodiment, handle  70  is formed with the cap as a single disposable unit. Other forms of attaching handle  70  to the vial cap include thermal bonding, adhesive bonding, bonding by chemical reaction, or other non-detachable permanent means. 
         [0030]    The uses of the terms “a” and “an” and “the” and similar references in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. 
         [0031]    While the invention has been illustrated and described in detail in the drawings and foregoing description, the same is to be considered as illustrative and not restrictive in character, it being understood that only the preferred embodiment has been shown and described and that all changes and modifications that come within the spirit of the invention are desired to be protected. In addition, all references cited herein are indicative of the level of skill in the art and are hereby incorporated by reference in their entirety. 
       LISTING OF CERTAIN EMBODIMENTS 
       [0032]    1. A tissue extraction vial comprising: 
         [0033]    a container having a mouth and a retrieval port, the retrieval port also having a closure configured to permit access into the container; 
         [0034]    a cap, and 
         [0035]    a blade configured to capture a tissue sample; 
         [0036]    wherein the blade is attached to the cap; 
         [0037]    wherein the cap is adapted to mate with the mouth, and; 
         [0038]    wherein the cap is adapted to sealingly mate with the mouth of the container. 
         [0039]    2. The tissue extraction vial of embodiment 1 wherein the blade defines an internal lumen. 
         [0040]    3. The tissue extraction vial of embodiment 2 wherein the blade is circular. 
         [0041]    4. The tissue extraction vial of embodiment 3 wherein the circular blade has a diameter between about 2 mm and about 4 mm. 
         [0042]    5. The tissue extraction vial as in any preceding embodiment wherein the blade is made of stainless steel. 
         [0043]    6. The tissue extraction vial as in any preceding embodiment wherein the blade has a height of about 5 mm. 
         [0044]    7. The tissue extraction vial of any preceding embodiment further comprising: 
         [0045]    a plunger, wherein the plunger is configured to dislodge the tissue sample. 
         [0046]    8. The tissue extraction vial as in any preceding embodiment wherein the closure is a septum that can be pierced by a needle. 
         [0047]    9. The tissue extraction vial as in any preceding embodiment further comprising a liquid medium inside the container. 
         [0048]    10. The tissue extraction vial as in embodiment 9 wherein the liquid medium is a solution. 
         [0049]    11. The tissue extraction vial as in embodiment 9 or 10 further comprising a chemical preservative capable of preserving the tissue sample inside the container. 
         [0050]    12. The tissue extraction vial as in embodiment 9 or 10 further comprising a preservative with a permeable cryoprotectant agent inside the container. 
         [0051]    13. The tissue extraction vial as in embodiment 9 or 10 further comprising one of the following preservatives inside the container: dimethyl solfoxide, ethylene glycol, propylene glycol, or glycerol. 
         [0052]    14. The tissue extraction vial as in embodiment 9 or 10 further comprising one of the following enzymes adapted to digest a tissue sample inside the container. 
         [0053]    15. The tissue extraction vial of any preceding embodiment wherein the retrieval port is at one end of the container, and the mouth is at another end of the container. 
         [0054]    16. A method of extracting tissue comprising the steps of: 
         [0055]    acquiring a tissue extraction device comprising:
       a container having a mouth and a retrieval port, the retrieval port also having a closure configured to permit access into the container;       
 
         [0057]    a cap, and 
         [0058]    a blade configured to capture a tissue sample; 
         [0059]    wherein the blade is attached to the cap; 
         [0060]    wherein the cap is adapted to mate with the mouth, and; 
         [0061]    wherein the cap is adapted to sealingly mate with the mouth of the container. 
         [0062]    obtaining a tissue sample using the blade; 
         [0063]    placing the tissue sample inside the container, and; 
         [0064]    mating the cap to the container mouth. 
         [0065]    17. The method of extracting tissue in embodiment 14 wherein the tissue extraction device has a liquid medium inside the container. 
         [0066]    18. The method of extracting tissue in embodiment 16 wherein the liquid medium is a solution. 
         [0067]    19. The method of extracting tissue in embodiment 16 or 17 wherein a chemical preservative adapted to preserve the tissue sample and cells is also inside the container. 
         [0068]    20. The method of extracting tissue in embodiment 16 or 17 wherein a preservative that includes a permeable cryoprotectant agent is also in the container. 
         [0069]    21. The method of extracting tissue in embodiment 16 or 17 wherein one of the following preservatives is also inside the container: dimethyl solfoxide, ethylene glycol, propylene glycol, or glycerol. 
         [0070]    22. The method of extracting tissue in embodiment 14 comprising the additional steps of: 
         [0071]    acquiring a tissue extraction device wherein the solution in the inner void incorporates enzymes adapted to digest the tissue sample to liberate cells from the tissue sample; 
         [0072]    extracting the cells through the retrieval port. 
         [0073]    23. A tissue extraction vial comprising: 
         [0074]    a container; 
         [0075]    a cap for the container; 
         [0076]    a blade configured to capture a tissue sample, the blade attached to the cap; and 
         [0077]    a needle-penetrable septum adapted to provide access into the container. 
         [0078]    24. The vial of embodiment 23 wherein the blade is receivable within the container when the cap is mated to the container. 
         [0079]    25. A cap assembly for capping a container, comprising: 
         [0080]    a cap matable with a container; 
         [0081]    a blade configured to capture a tissue sample, the blade attached to the cap; and 
         [0082]    a needle-penetrable septum mounted in the cap and adapted to provide access through the cap into the container when the cap is mated to the container. 
         [0083]    26. The cap assembly of embodiment 25, wherein the blade defines an inner lumen having a longitudinal axis, and wherein at least a portion of the septum is aligned with said longitudinal axis.