DEVICE AND ASSEMBLIES FOR ORIENTED TRANSPORT, MICROSCOPIC INVESTIGATION AND ORIENTED EJECTION OF A TISSUE GRAFT OR IMPLANT

Abstract

The invention provides a device for secure support, storage and/or transport of a tissue graft or an implant, which allows for precise microscopic investigation and/or evaluation and quality control of the tissue graft or implant during processing as well as prior to introduction into a living body. The main body of the device is a cannula or a cartridge comprising rectangular shape, preferably with rounded edges, having two opposite openings of different sizes and shapes. The first opening has a round shape with a diameter suitable for connection to a tube or a syringe nozzle. The second opening has an elliptical, round, biconvex or rectangle shape, in the latter case preferably with rounded edges that is small enough to be inserted into a small surgical incision. The transparency and preferably rounded, rectangular shape of the main body of the device permits microscopic examination of the tissue graft or implant within the device, as well as secures the tissue graft or implant position during delivery and implantation. The elliptical, round, biconvex or preferably rounded rectangle inner shape of the second opening ensures the correct positioning of the tissue graft or implant during surgical implantation, thereby preventing issues such as loss of the tissue graft or implant orientation. The invention further provides assemblies for loading, storing and/or transporting a tissue graft or implant comprising the device according to the invention. The invention further provides a washing assembly and a method for preparing a tissue graft or implant.

Claims

1. Device (10) for secure support storage and/or transport of a tissue graft or implant (11) for ophthalmological interventions, comprising: a first opening (1), a main body (2), a taper area (3), and a second opening (4), characterized in that the first opening (1) has a round shape and a funnel-like design configured to connect with a tube (5) or a syringe (8, 12) and the main body (2) is transparent and has a rectangle shape and the taper area (3) is transparent and has a an elliptical, round, biconvex or rectangle shape, and wherein the main body (2) comprises at least two flat and parallel opposite sites.

2. Device (10) according to claim 1, wherein the second opening (4) has an elliptical, round, biconvex or rounded rectangle shape.

3. Device (10) according to claim 1, wherein the outer wall of the second opening (4) has a different shape than the inner wall.

4. Device (10) according to claim 1, wherein the outer wall of the taper area (3) has a different shape than the inner wall.

5. Device (10) according to claim 1, wherein the device (10) consists of glass, preferably borate glass or consists of plastic, wherein said plastic is transparent and has a refractive index (r.sub.i) in the range of r.sub.i=1.30 to r.sub.i=1.71, preferably r.sub.i=1.30 to r.sub.i=1.65, most preferably 1.30 to r.sub.i=1.60, which is similar to the r.sub.i=1.33 to 1.34 (at 20° C. and 600 nm) of balanced salt solution (BSS), such as Hank's BSS, Earle's BSS, Tyrode's salt solution, Alsever's salt solution, phosphate-buffered saline (PBS), Tris-buffered saline (TBS), Puck's salt solution, Gey's salt solution, Ringer's salt solution, Simm's salt solution and related buffered saline solutions.

6. Device (10) according to claim 5, wherein the inner surface of the device (10) comprises a hydrophobic coating, selected from coatings comprising acrylate, organo-siloxane, silane, epoxy, a polymer, Molybdenum disulfide, Molybdenum disulfide/graphite, Tungsten disulfide or graphite, preferably selected from coatings comprising a polymer, organo-siloxane, silane, acrylate or epoxy; or a hydrophilic coating, selected from coatings comprising any hydrophilic polymer/hydrogel, preferably selected from coatings comprising poly(ethylene glycol), poly(acrylate), poly(methacrylate) or a UV/photo-active polymer; and/or a surface patterning, being a micro- or nanostructured surface pattern or a combination thereof (e.g. nano-microstructuring) in the range of 100 nm to 20000 nm, preferably between 300 nm to 5000 nm and more preferably between 500 nm to 2500 nm.

7. Device (10) according to claim 1, further comprising a cap (13, 14, 15) for the first opening (1) and/or a cap (13, 14, 15) for the second opening (4).

8. Assembly for loading, storage and transport of a tissue graft or implant (11), comprising the device (10) according to claim 1, at least one syringe (8, 12), a tube (5), and at least one cap (13, 14, 15).

9. Assembly according to claim 8, further comprising a transport device.

10. Assembly according to claim 9, wherein the transport device is a device holder (20) comprising a round neck (18) with a holder (19), finger tips which are attached on an oval shaped platform with a cavity (16), and a tilted stripe (17), which is suitable to be positioned inside a container, such as a standard tissue culture flask or a tissue culture flask with two openings (40).

11. Assembly according to claim 10, wherein the finger tips and the oval shaped platform with a cavity (16) do not optically interfere with the main body (2) and the taper area (3) of the device (10).

12. Washing assembly for washing and staining a tissue graft or implant (11), comprising a macroporous material (21) with interconnected pores and a porosity between 10 μm and 600 μm, the device (10) according to claim 1, at least one syringe (8, 12), and a 2-way extension line (31) or a 3-way stopcock (32).

13. Method for preparing a tissue graft or implant (11) using the device (10) according to claim 1, comprising the steps: a) providing a tissue graft or implant (11), b) loading the tissue graft or implant (11) into the device (10), c) sealing the device (10) by at least one cap (13, 14, 15), d) evaluation and quality control of the tissue graft or implant (11) inside the device (10), e) transport of the device (10) with the tissue graft or implant (11), f) optionally evaluation and quality control of the tissue graft or implant (11) inside the device (10), g) washing and staining of the tissue graft or implant (11) inside the device (10).

14. Method according to claim 13, wherein the device (10) is transported within the device holder (20).

15. Method according to claim 13, wherein evaluation and quality control of the tissue graft or implant (11) is performed with the device (10) containing the tissue graft or implant (11) being inside the transport device and inside a container, such as a standard tissue culture flask or a tissue culture flask with two openings (40).

Description

FIGURES

[0245] In the following, the invention is additionally illustrated in further detail by 11 figures, wherein

[0246] FIG. 1: shows different designs of the device,

[0247] FIG. 2: show a cross section of the device,

[0248] FIG. 3: shows parts of the assembly for loading, storage and/or transport of the tissue graft or implant,

[0249] FIG. 4: illustrates the process of tissue graft or implant loading into a device,

[0250] FIG. 5: shows different tissue graft or implant positioning inside the device,

[0251] FIG. 6: illustrates a preparation of device loaded with a tissue graft or implant aiming for direct introduction into a living body (short-term transport assembly),

[0252] FIG. 7: illustrates different caps for closing the first and second opening of the device,

[0253] FIG. 8: shows a possible device holder design from different perspectives, as well as a tissue culture flask with two openings,

[0254] FIG. 9: illustrates the application of a macroporous material as part of the assembly for washing and/or staining of a tissue graft or implant,

[0255] FIG. 10: illustrates the release of a tissue graft or implant from the device,

[0256] FIG. 11: illustrates application of a 2-way extension line or a 3-way stopcock of the washing assembly for washing and/or staining of a tissue graft or implant.

[0257] FIG. 1 illustrates three different possible designs (labelled A, B and C) of the device according to the invention. Every design is illustrated from the outside, view from the front and drawn transparently, so that the inner walls can be seen and the dimensions of some of the inner and outer compartments are illustrated. In all designs, the first opening (1), the main body (2), the taper area (3) and the second opening (4) are shown. The dimensions of the outer walls of the main body are marked with the letter (a) and are closer described by the letters (d) and (e) in the front view, the dimensions of the outer walls of the taper area are marked with the letter (b) and are closer described by the letters (f) and (g) in the front view. For all designs showed, the main body (2) has the same dimensions and shape, but can also be different according to the invention. The dimensions of the inner walls of the taper area are marked with the letter (c) and are closer described by the letter (h) and (i) in the front view. The following table gives an overview about the used markings.

TABLE-US-00004 Main body (2) d Outside distance between the side walls (“width”) e Outside distance between the two flat and parallel opposite sides (“height”) Taper area (3) f Outside distance between the side walls (“width”) g Outside distance between the top and bottom walls (“height”) h Distance between outer wall and inner wall at the smallest point i Radius of the curvature of the corners of the inner wall

[0258] Design A shows a second opening (4) which inner wall has a rounded rectangle shape with two flat opposite walls, while the outer wall has an oval shape. The figure illustrates clearly the flat and parallel opposite sides of the main body (2), whereby the main body (2) has the following dimensions d=3.7 mm and e=2.7 mm. Due to the oval outer shape, the taper area (3) is slightly smaller compared to the main body (2) to fit better in a surgical incision as described above. The taper area (3) of the device (10), design A shown in FIG. 1 has the following dimensions: f=3.7 mm and g=2.6 mm. The inner hollow compartment of the taper area (3) has rounded rectangle shape which is closer described by the following dimensions h=0.3 mm and i=0.7 mm.

[0259] Design B shows a second opening (4) which inner and outer wall has an oval shape. The main body (2) with its flat and parallel opposite walls has the same dimensions (d) and (e) as for the design A. The dimensions of the taper area are: f=3.4 mm, g=2.5 mm and h=0.3 mm. The use of an oval shape also of the inner wall of the taper area has the advantage of a smoother tissue graft or implant ejection through the second opening of device design B, in comparison to design A.

[0260] Design C illustrates another device (10) according to the invention. The main body (2) with its flat and parallel opposite walls has the same dimensions (d) and (e) as for the design A. The taper area (3) does not have a constant thickness as design A and B and is linearly decreasing, until it remains in a constant thickness towards the second opening. At the smallest section, the taper area has the following dimensions: f=2.5 mm, g=1.8 mm and h=0.3 mm and the inner walls as well as the outer walls of the taper area (3) have a rounded rectangle shape. The second opening (4) has a rounded rectangle shape as well.

[0261] All designs allow for tissue graft or implant injection in small surgical incisions (2.4 mm to 3.0 mm incision width) as described above.

[0262] FIG. 2 shows a cross section of the device (10). Dimensions of the inner hollow compartments of the main body (2) and the taper area (3) are marked by the letters (m) and (n). Additionally, the dimension of the inner hollow compartment of the first opening (1) is marked by the letter (k). According to the invention, the first opening (1) can have a dimension (k) between 3 mm and 6 mm. The main body (2) can have a dimension (m) between 1 mm and 5 mm and the taper area (3) can have a dimension (n) between 0.8 mm and 2 mm. The overall length (l) of the device (10) is between 25 mm and 50 mm.

[0263] FIGS. 3 (A) to (E) show parts of the assembly for loading, storage and transport of a tissue graft or implant. FIG. 3 (A) illustrates a tube (5) included in one embodiment of the invention with a Luer Slip connector (6). The connection of a tube (5) with a syringe (8) via a Luer Lock connector (7) is illustrated in FIG. 3 (B). The second opening (4) of the device (10) is connected to the tube (5) as can be seen in FIGS. 3 (C) and (D). For loading a tissue graft or implant (11) into the device (10), the second opening (4) of the device (10) is connected to a tube (5) and the tube (5) is connected to a syringe (8) for example via a Luer Slip connector (6) (FIG. 3 (E)).

[0264] FIGS. 4 (A) to (C) illustrate the process of tissue graft or implant loading into a device. For loading the tissue graft or implant (11) into the device (10), a syringe (8) which is for example preferably filled with balanced salt solution or liquid nutrition medium, is connected to the second opening (4) of the device (10) by attaching one end of a tube (5) to the second opening (4) and the other end of the tube (5) to the syringe (8) via a Luer Lock connector (7). Subsequently, the syringe plunger is moved to push out excess air from the device (10) and from the tube (5), there should be no air bubbles left in the device (10) and in the tube (5) (FIG. 4 (A)).

[0265] For loading the tissue graft or implant (11) into the device (10), the device (10) is attached to a laboratory dish/petri dish (9) containing the tissue graft or implant (11) in a suitable liquid medium (e.g. BSS or nutrition medium). Loading is performed from the first opening (1) which is big enough for gentle tissue graft or implant (11) uptake due to its funnel-like design (the diameter of the opening increases slightly towards the edge). Therefore, the first opening (1) of the device (10) is gently placed on top of the tissue graft or implant (11) without touching the tissue graft or implant (11), and the syringe plunger is moved to upload the tissue graft or implant (11) inside the device (10) by use of hydrodynamic flow (FIGS. 4 (B) and (C)).

[0266] For storage and/or transport and evaluation, the tissue graft or implant (11) is positioned inside the main body (2) of the device (10), which can be called ‘transport position’ (FIG. 4 (C) and FIG. 5 (A)). For direct introduction of the tissue graft or implant (11) into for example a living body, the tissue graft or implant (11) can be positioned partially inside the taper area (3) of the device (10), which can be called ‘injection position’ (FIG. 5 (B)). In this position, the tissue graft or implant (11) is partially rolled in/slightly compressed, which ensures stable positioning inside the device (10), e.g. for the attachment of a BSS-filled syringe (8 or 12) at the first opening (1) of the device (10) (FIG. 6 (A)). The tissue graft or implant (11) could be positioned also completely inside the taper area (3) for the ‘injection position’. However this increases the probability of unwanted slipping out of the tissue graft or implant (11) from the device (10) during attachment of a syringe (8 or 12) at the first opening (1) of the device (10) since in this case there is not much space left towards the second opening (4) of the device (10).

[0267] FIGS. 6 (A) to (C) illustrate the preparation of the device (10) loaded with a tissue graft or implant (11) aiming for direct introduction into a living body. After loading the tissue graft or implant (11) into the device (10), a second liquid-filled (e.g. balanced salt solution or nutrition medium) syringe (12) is directly connected to the first opening (1) of the device (10). At this point, the tube (5) connecting the second opening (4) and the syringe (8) used for loading the device (10) are still connected (FIG. 6 (A)). After connecting the second syringe (12) to the first opening (FIG. 6 (B)), the tube (5) with the syringe (8) is disconnected (FIG. 6 (C)) and the device is ready for ejection of the tissue graft or implant (11), for example for injection into a living body.

[0268] According to the invention, different kind of caps are suitable to close the first (1) and the second opening (4) of the device (10). FIGS. 7 (A) to (D) illustrate some suitable caps. A first cap (13, 15) closes the first opening (1) and a second cap (14) closes the second opening (4) as shown in FIGS. 7 (A) and (C). In both embodiments, the main body (2) is left uncovered and can be used for microscopic examination and/or evaluation of the tissue graft or implant (11) by eye. At least one of the used caps is permeable to allow for exchange of oxygen and nutrients during transport and/or storage of the tissue graft or implant (11) inside the device (10). Further embodiments of the invention are shown in FIGS. 7 (B) and (D). The first opening (1) is still closed by a first cap (13, 15) and the second opening (4) is closed by a Luer Slip connector (6) equipped with a short flexible tubing (5), which can be of the same material as described for the tubing (5) for tissue loading (FIG. 3). These embodiments have the advantage that not only the main body (2) is left uncovered but also the taper area (3), therefore both parts of the device (10) can be used for microscopic examination and/or evaluation of the tissue graft or implant (11) by eye.

[0269] For secure storage and/or transportation of the device (10) with the tissue graft or implant (11) the present invention further provides a device holder (20) as shown in FIG. 8 (A). The device holder comprises finger tips with an oval holder with a cavity (16), wherein the finger tips have cavities with a shape matching the side of the device (10) for easy assembly and gentle device (10) removal as well as for holding the device (10) tightly. Furthermore, the shape of the finger tips with an oval holder with a cavity (16) secures the orientation/position of the device (10) during storage and/or transportation. Especially, this prevents the device (10) from spinning. The finger tips with the oval holder with a cavity (16) do not interfere with the main body (2) and the taper area (3) of the device (10), which therefore can still be used as microscope examination area. The finger tips with the oval holder with a cavity (16) are attached to a tilted stripe (17) which is attached to a round neck (18) which from one side allows the insertion into a standard tissue culture flask or a tissue culture flask with two openings (40), but from another side holds tight to the neck of the neck part of the standard tissue culture flask or to one neck of the tissue culture flask with two openings (40). This prevents the movements of the whole transport device inside the standard tissue culture flask or inside the tissue culture flask with two openings (40). The round neck (18) has a holder (19) in the center which allows its manipulation with forceps and/or the end user's fingers, for secure and easy insertion and removal of the device holder (20) from the standard tissue culture flask or from the tissue culture flask with two openings (40). The overall dimensions of the device holder (20) are made in a way that the one end of the tilted stripe (17) touches the bottom of the standard tissue culture flask or the bottom of the tissue culture flask with two openings (40) while the other end with the round neck (18) is in the neck of the standard tissue culture flask or in one neck of the tissue culture flask with two openings (40) touching the closing cap of the standard tissue culture flask or one closing cap of the tissue culture flask with two openings (40) when the standard tissue culture flask or the tissue culture flask with two openings (40) is closed. FIG. 8 (B) shows a tissue culture flask with two openings (40), suitable for insertion of the device holder (20), including the device (10). The design of the tissue culture flask with two openings (40) allows for easy aspiration of the medium (e.g. for fungal tests) directly in the device (10) from one opening (43), while the other opening (44) can be used to insert the device holder (20) including the device (10). A screw cap (41, 42) at both necks (43, 44) ensures a tight and leak-proof sealing of the tissue culture flask with two openings (40) during storage and/or transport and evaluation of the tissue graft or implant (11) being inside the device (10), which is preferably kept in place with the device holder (20) inside the tissue culture flask with two openings (40).

[0270] The use of a macroporous material (21) for washing and staining is shown in FIGS. 9 (A) to (F). The first opening (1) of device (10) is connected to a syringe (12) and the second opening (4) of the device (10) is positioned on top of the macroporous material (21) (FIGS. 9 (A) and (B)). In one embodiment, the device (10) is pressed closely against the macroporous material (21) (FIGS. 9 (C) and (D)), thereby providing a tight connection between macroporous material (21) and second opening (4). In another embodiment, the device (10) is pressed against the macroporous material (21) in a way that the second opening (4) as well as the taper area (3) are enclosed by the macroporous material (21) (FIGS. 9 (E) and (F)), therefore providing an even more tight connection. The macroporous material (21) has such dimensions that it is big enough to uptake the solution pressed out of the device (like a sponge) and that the device (10) does not break through at the bottom at macroporous material (21).

[0271] FIGS. 10 (A) to (C) illustrate the release of the tissue graft or implant (11) out of the device (10) by pushing a syringe (8 or 12) connected to the first opening (1). The syringe (8 or 12) is not shown. The tissue graft or implant is gently moved by a hydrodynamic flow in an oriented manner. Firstly, the tissue graft or implant (11) is positioned inside the taper area (3) of the device (10) (‘injection position’, FIG. 10 (A)). By pushing the plunger of the syringe (8 or 12), the tissue graft or implant (11) is slipping out of the second opening (4) of the device (10) (FIG. 10 (B)) and finally leaves the device (10) (FIG. 10 (C)).

[0272] Additionally, the washing assembly comprises either a 2-way extension line (31) or 3-way stopcock (32), both comprising flexible tubes (33, 34) with male (preferably Luer Lock or Luer Slip) connectors (26), as shown in FIG. 11. In one embodiment of the invention, the washing assembly further comprises tube clamps (27, 28) and closing caps (22, 23) for the female endings (24, 25) of the 2-way extension line (31) or 3-way stopcock (32). The tube clamps (27, 28) can be used to interrupt the flow inside a tube (29, 30) used in the washing assembly.

[0273] The 2-way extension line (31) as well as the 3-way stopcock (32) have a flexible tube (34, 33) with a male connection (26) for attaching the device (10). The male connection (26) is preferably a Luer Lock or a Luer Slip connector. Both the 2-way extension line (31) and the 3-way stopcock (32) have two other connectors, which are preferably female connectors (24, 25). The female connectors (24, 25) are preferably Luer Lock or Luer Slip connectors. The female connectors (24, 25) are suitable to connect a conventional syringe (8) or (12) and can optionally be closed with a cap (22, 23). As shown in FIG. 11, the connections with the female connectors (24, 25) are in the case of the 2-way extension line (31) equipped with tubes (29, 30), while the 3-way stopcock (32) has no tubes connected to the female connectors (24, 25). However, the availability of tubes (29, 30) is not a must.

[0274] For tissue graft or implant (11) washing and staining, a syringe (8) filled with balanced salt solution (BSS) is attached to one female connector (24) of the 2-way extension line (31) or the 3-way stopcock (32) and another syringe (12) filled with staining solution (e.g. trypan blue) is attached to the other female connector (25). The volume of the BSS-filled syringe (8) is preferably greater than the volume for the second staining solution-filled syringe (12). The syringes (8 or 12) are not shown in FIG. 11.

[0275] The use of the assemblies shown in FIG. 11 is described above in detail.

[0276] For removing air from the assembly, in the case of using a 2-way extension line (31), a clamp (28) of the tube (30) at the position (25) with the staining solution-filled syringe (12) remains open, while another clamp (27) at the BSS-filled syringe position (24) outlet is fully closing the tube (29). For the 3-way stopcock (32), the tap position is adjusted in a way that the staining solution-filled syringe (12) at position (25) and the male connection (26) outlet are connected while the position (24) with the BSS-filled syringe (8) remains closed.

[0277] First, the tubing (30) and the position (25) attached to the staining solution-filled syringe (12) is fully flushed with staining solution. The staining solution must not enter the male connection (26) of the assembly (31, 32), since this step is only needed for air removal.

[0278] Next, complete air removal and flush of the assembly with BSS solution needs to be performed. In the case of using a 2-way extension line (31), a clamp (28) of the tube (30) at the position (25) with the staining solution-filled syringe (12) is fully closing the tube (30), while another clamp (27) of the tube (29) at the BSS-filled syringe (8) position (24) outlet remains open. For the 3-way stopcock (32), the tap position is adjusted in a way that the BSS-filled syringe (8) at position (24) and the male connection (26) outlet are connected while the position (25) with the staining solution-filled syringe (12) remains closed.

[0279] Now, the assembly is fully flushed with BSS by pushing the plunger of the BSS-filled syringe (8). There should be no air left in the assembly as well as there should be no staining solution eluting from the male connection of the assembly. Fourth, the device (10) containing the tissue graft or implant (11) is attached via the first opening (1) to the male connector (26) of the 2-way extension line (31) or 3-way stopcock (32) and placed into a BSS-filled laboratory dish/petri dish (9, not shown in FIG. 11) to prevent air infiltration to the device (10).

[0280] To decrease the probability of tissue graft or implant (11) slipping out of the device (10), the second opening (4) of the device (10) can be closed by a cap (13, 14, 15) or can be placed onto/into the macroporous material (21) from the washing assembly during the whole staining and washing process. If applicable, the liquid transport medium can now be flushed out of the device (10) prior staining by gently pushing the plunger of the BSS-filled syringe (8) at position (24). Alternatively, staining of the tissue graft or implant (11) can be performed without prior washing with BSS solution.

[0281] Next, for the use of a 2-way extension line (31), the clamp (27) at the BSS-filled syringe (8) position (24) is fully closing the tube (29), while the other clamp (28) of the tube (30) at the position (25) with the staining solution-filled syringe (12) outlet remains open. In case of the use of a 3-way stopcock (32), the tap position is adjusted in a way that the staining solution-filled syringe (12) at position (25) and the male connection (26) outlet are connected while the position (24) with the BSS-filled syringe (8) remains closed.

[0282] Now the staining solution is gently added to the tissue graft or implant (11) in the device (10) by pushing the plunger of the staining solution-filled syringe (12) at position (25) and left for at least 1-2 minutes for sufficient staining. Extended staining will cause increased cell damage. After staining, in the case of using a 2-way extension line (31), the clamp (28) at the position (25) with the staining solution-filled syringe (12) is fully closing the tube (30), while the other clamp (27) of the tube (29) at the BSS-filled syringe (8) position (24) outlet remains open. For the 3-way stopcock (32), the tap position is adjusted in a way that the BSS-filled syringe (8) at position (24) and the male connection (26) outlet are connected while the position (25) with the staining solution-filled syringe (12) remains closed.

[0283] Lastly, the device (10) is fully flushed with BSS by pushing the plunger of the BSS-filled syringe (8). The solution in the device (10) should be completely transparent and no staining solution should remain. Now, the device (10) and the respective tissue graft or implant (11) are ready for ejection of the tissue graft or implant (11), especially for injection the tissue graft or implant (11) into the living body.

LIST OF REFERENCE NUMERALS

[0284] 1 First opening [0285] 2 Main body [0286] 3 Taper area [0287] 4 Second opening [0288] 5 Tube [0289] 6 Luer Slip connector [0290] 7 Luer Lock connector [0291] 8 Syringe [0292] 9 Laboratory dish/petri dish [0293] 10 Device [0294] 11 Tissue graft or implant [0295] 12 Syringe [0296] 13 Cap [0297] 14 Cap [0298] 15 Cap [0299] 16 Finger tips with oval shaped platform with a cavity [0300] 17 Tilted stripe [0301] 18 Round neck [0302] 19 Holder [0303] 20 Device holder [0304] 21 Macroporous material [0305] 22 Cap [0306] 23 Cap [0307] 24 Female connector [0308] 25 Female connector [0309] 26 Male connector [0310] 27 Tube clamp [0311] 28 Tube clamp [0312] 29 Tube [0313] 30 Tube [0314] 31 2-way extension line [0315] 32 3-way stopcock [0316] 33 Tube [0317] 34 Tube [0318] 40 Tissue culture flask with two openings [0319] 41 Screw Cap [0320] 42 Screw Cap [0321] 43 Neck with thread [0322] 44 Neck with thread

REFERENCES

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