Cell tissue culture and transport box

Abstract

A cell tissue culture and transport dish comprising: a dish body having an accessible interior and a top opening; a filter disposed within the dish body and communicating the interior of the dish body with an exterior of the dish body, the filter being configured to permit gas exchange between the interior and exterior of the dish body while preventing contamination of the cell tissue culture; a closure means for closing the top opening of the dish body, the closure means being fluid- and gas-tight; and at least one aseptic fluid connection through the dish body, said at least one aseptic fluid connection including two fluid inlet connections and one fluid outlet connection. Abstract figure: Figure 1

Description

Title of the invention: Cell tissue culture and transport box technical field

[0001] The present disclosure falls within the domain of cell tissue culture and transport dishes. Previous technique

[0002] The process of manufacturing cellular tissue, such as artificial skin, involves the use of a sterile environment and a plurality of culture media introduced sequentially. The process can be time-consuming and involve many manipulations of the closed system.

[0003] Typically, stem cells are placed in a culture dish. This culture dish constitutes a closed environment in which the different layers of the skin will be created through the addition of culture media. Each time a medium needs to be added to the culture, the operator uncaps / unscrews / opens the dish and introduces the culture medium. Then, they recap / screw / close the dish, set it down, and let it rest. They then take the dish, uncap it, introduce the medium, and recap it in this way for each culture medium.

[0004] On the one hand, the multiple opening and closing operations of the dish can lead to contamination of the culture medium. On the other hand, this requires a significant amount of operator time, which does not allow for large-scale multiplication of cell tissue culture. Summary

[0005] A cell tissue culture and transport box is proposed comprising: a box body having an accessible interior and top opening; a filter disposed in the box body and communicating the interior of the box body with an exterior of the box body, the filter being configured to permit gas exchange between the interior and exterior of the box body, while preventing contamination of the cell tissue culture; a closure means closing the top opening of the box body, the closure means being fluid and gas tight; and at least one aseptic fluid connection through the box body, said at least one aseptic fluid connection including two fluid inlet connections and one fluid outlet connection.

[0006] The features described in the following paragraphs may optionally be implemented independently of each other or in combination with each other: - the closing mechanism is airtight against fluids and gases - The closure device is for single use only, so that once detached from the top opening, the inside of the box body is accessible through the top opening. - The closure device is a lid or a cap. - The closure means is sealed, screwed or clipped to the box body, for example by an O-ring, heat sealing or ultrasound - the closure mechanism is transparent - the closing means retains its mechanical properties up to an exposure temperature of approximately -200°C. - the closure means is made of polyethylene terephthalate or polyethylene. - the filter is positioned at the level of said at least one fluidic connection. - the filter is higher than the culture medium to ensure its function of gas exchange and not to be wetted by the culture medium. - the filter is arranged in a horizontal or inclined part of the inside of the box body - the inclined part being inclined towards the insert. - the filter is positioned vertically below an area for gripping the insert. - the filter is arranged in the distal box body of a culture zone of the insert. - the filter is made of polyethylene, polyurethane, or polypropylene. - the box also includes a removable insert suitable for placement inside the body of the box, the insert being suitable for receiving a cell tissue culture. - The insert is removable from the box body via the top opening. - the insert has a culture zone and a gripping zone connected to the culture zone, the gripping zone being adapted to manipulate the insert relative to the box body. - The culture area includes a top opening suitable for accommodating cell tissue culture. - the growing area includes a porous base. -the porous base is opposite the top opening when the insert is placed in the box body. - The porous base of the insert is made of polyester, polyethylene or polycarbonate - the pore size of the porous base is 0.2 to 4 µm, preferably 0.2 to 0.6 µm / - the density of the pores of the porous base is from l*106 to 5*106, preferably from l*106 to 3*106 pores per cm2. - the gripping area is inclined towards the growing area - The growing area of ​​the insert is rounded in shape - the culture zone of the insert is a ring - the culture zone of the insert has a thickness corresponding to the thickness of a working surface of the box body adapted to receive the culture zone of the insert. - the culture zone of the insert has a shape corresponding to a shape of a working surface of the box body adapted to receive the culture zone of the insert. - said insert is resistant to dimethyl sulfoxide (DMSO). - said insert includes at least one connection adapted to cooperate removably with the inside of the box body. - said at least one connection is at least one tab adapted to be inserted removably into a housing inside the box body. - the dwelling is an open recess. - the insert is made of polypropylene, fluorinated ethylene propylene, polycarbonate. - the interior of the box body includes a working area suitable for receiving cell tissue culture and fluid culture media allowing the cell tissue to grow. - The work area includes a work surface adapted to accommodate the culture zone of the insert. - the work surface being preferably flat. - the work surface has undergone plasma surface treatment. - the inside of the box body includes a part inclined towards the work surface. - the work surface is transparent. - the box body is substantially parallelepiped and has rounded edges. - the work surface is removable from the rest of the box body. - the box body is made of polyester, polypropylene, fluorinated ethylene propylene, polycarbonate, polycarbonate alloy or Eastman Tritan™ Copolyester MP100. - the two fluidic connections for fluid inlet are arranged vertically above the culture area. - The fluid outlet connection is arranged vertically at the same level as, or below, the culture zone. - The two fluid inlet connections are ports with a tubular extension extending into the box body. - the tubular extension has one end positioned vertically above the insert. - one end of the fluidic inlet connections inside the box body is beveled. - the bevel is oriented towards the upper opening. - the bevel is approximately 45 degrees. - the tubular extension is sealed to the box body by welding and the bevels obtained using an aseptic disconnecting machine. - the fluid outlet connection is a septum, a hose clamp, a barbed fitting associated with a radial crimping system or a Luer-Lock type conical fitting. - said at least one fluidic connection is resistant to dimethyl sulfoxide (DMSO). Brief description of the drawings

[0007] Other features, details and advantages will become apparent upon reading the detailed description below and analyzing the accompanying drawings, in which: Fig. 1

[0008] Fig. 1 is a schematic perspective view from the top of a cell tissue culture and transport box according to one embodiment; Fig. 2

[0009] Fig. 2 is a schematic perspective view of the underside of the box in Fig. 1. Fig. 3

[0010] Fig. 3 is another schematic perspective view of the top of the box in Fig. 1; Fig. 4

[0011] The [Fig.4] is a schematic cross-sectional view of the box of the [Fig.1];

[0012] ; Fig. 5

[0013] Figure 5 is a schematic perspective view of an insert for the box in Figure 1; and Fig. 6

[0014] Fig. 6 is a schematic view of an embodiment of the insert where a porous base of the insert is detachable; Fig. 7

[0015] Figure 7 is a schematic view of another embodiment of the insert where a The porous base of the insert is detachable; Fig. 8

[0016] Figure 8 is a schematic view of an embodiment of the box where the box has a removable bottom; and Fig. 9

[0017] Figure 9 is a schematic view of another embodiment of the box where a The filter is arranged vertically. Description of the implementation methods

[0018] With reference to Figures 1 to 4, a cell culture and transport (CCT) dish 10 according to one embodiment will be described. The dish 10 comprises a body 12 having an accessible interior 14 and a top opening 16. The top opening 16 is closed by a closure means 18. The closure means 18 is preferably fluid- and gas-tight. The dish 10 includes at least one aseptic fluid connection 20 through the body of the box 12. In the example of [Fig. 1], the box 10 includes three aseptic fluid connections 20: two inlet fluid connections 22 for culture fluids, and one outlet fluid connection 24 for fluids. The inlet fluid connections 22 allow the delivery of fluids, preferably liquids, for the culture of TC cell tissue, while the outlet fluid connection 24 allows the TC cell tissue culture to be drained of these same fluids. The interior 14 of the body of the box 12 includes a working area (or volume) 36 adapted to receive the TC cell tissue culture and the culture fluid media that allow the TC cell tissue to grow.

[0019] The box 10 further includes a filter 26 disposed in the box body 12 and communicates the inside of the box body 12 with an outside 28 of the box body 12. The filter 26 is configured to allow gas exchange between the inside 14 and the outside 28 of the box body 21, while preventing the passage of liquids.

[0020] The box 10 may optionally include a removable insert 30 disposed in the working area 36 and adapted to receive the TC cell tissue culture. The insert 30 can be inserted into the interior 14 of the box body 12 through the upper opening 16. Therefore, the upper opening 16 is sized to allow the passage of the insert 30. The insert 30 may have a culture area 34 adapted to receive and culture the TC cell tissue and a gripping area 35 allowing the insert 30 to be manipulated, in particular to be placed in and / or removed from the box body 12. The insert 30 will be described in more detail below, in relation to [Fig. 5]. According to other embodiments, the box 10 could not have an insert, so that the TC cell tissue culture is done directly on the inside 14 of the box body 12 at the level of the working area 36.

[0021] The working zone 36 is sized to contain an adequate volume of culture liquid. In the case of the insert 30, the working zone 36 has a slightly larger volume than the culture zone 34 of the insert 30.

[0022] The working area 36 of the body of the plate 12 includes a working surface 32 adapted to receive the TC cell tissue culture. In the case of plates with an insert, the working surface 32 is adapted to receive the culture area 34 of the insert 30. In the case of plates 10 without an insert, the TC cell tissue will be placed directly on the working surface 32 of the plate 10. In this latter case, the working surface 32 may have undergone a plasma surface treatment to allow the TC cell tissue to adhere to the working surface 32.

[0023] Whether with or without an insert, the working surface 32 can, for example, be transparent to allow visual monitoring of the TC cell tissue culture without having to open the closure means 18. The working surface 32 can be part The body of the box 12 delimits the interior 14 from the exterior 28. More broadly, all or part of the body of the box 12 could be transparent. The working surface 32 is preferably flat. It can be a reinforcement of the body of the box 12, as illustrated in the figures. The working surface 32 can be detached from the body of the box 12 so as to extract the TC cellular tissue once it has formed, as will be explained in relation to Figures 6 to 8.

[0024] The closure means 18 is used to create a closed system within the dish 10 that allows for the culture of TC cell tissue. The closure means 18 can be sealed to the dish body 12, for example by heat sealing or ultrasound. The closure means 18 may include a tab 19 that allows the closure means 18 to be removed manually without the use of instruments, such as a scalpel.

[0025] The closure means 18 can be single-use such that, once detached from the upper opening, the interior 14 of the box body 12 is accessible through the upper opening 16. Thus, the cell tissue culture and transport box 10 can be a single-use product. In the example shown in the figures, the closure means 18 is transparent to allow visual inspection of the cell tissue culture. The closure means 18 could also be opaque, partially transparent, colored, or clear. To withstand cryogenic conditions during the transport and storage of the cell tissue culture, the closure means 18 is preferably adapted to retain its mechanical properties down to an exposure temperature of approximately -200°C. The closure means 18 is, for example, made of polyethylene terephthalate or polyethylene.

[0026] The box body 12 can be made by plastic injection molding. The box body 12 can be made, for example, of polypropylene, fluorinated ethylene propylene, polycarbonate, or Eastman Tritan™ Copolyester MP100. The box body 12 can be adapted to retain its mechanical properties down to an exposure temperature of approximately -200°C, in order to allow for cryogenic storage of the box 10. The box body 12 can be resistant to dimethyl sulfoxide (DMSO). The box body 12 can be substantially parallelepiped-shaped with rounded edges to facilitate transport without wasting space. The box body 12 can generally have an ovoid shape when viewed from above.

[0027] The box body 12 can thus include a reinforcement 13 to receive one end of the gripping area 35 of the insert 30 so that the insert 30 does not protrude from the box 10.

[0028] The interior 14 of the box body 12 may include a portion inclined 38 towards the working surface 32. The inclined portion 38 may allow the flow of fluids towards the cell tissue culture area 36. The inclination of the inclined portion 38 is with reference to a horizontal plane H, the horizontal plane H containing the working surface. 32. The inclined part 38 is according to an embodiment with respect to the fluidic connections 20.

[0029] The inclined portion 38 can receive the filter 26. The filter 26 can be arranged horizontally in the inclined portion 38 or inclined, for example, at the same angle as the inclined portion as illustrated in Figures 1 to 4. The filter 26 could be arranged vertically in the inclined portion 38 as illustrated by the filter 26' in [Fig. 9]. An evaluation channel C could then extend from the inclined portion 38 through the filter 26' and out of the box. According to another embodiment, the filter 26 is positioned at the fluidic connections 20. The filter 26 can be positioned in the box body 12 distal to the working surface 32. This helps to limit the risk of wetting the filter 26. According to one embodiment, and as shown in the figures, the filter 26 can be positioned vertically below the gripping area 35 of the insert 30.Filter 26 can be made of polyethylene, polyurethane, or polypropylene. To withstand cryogenic conditions during the transport and storage of TC cell culture, filter 26 is preferably adapted to retain its mechanical properties down to an exposure temperature of approximately -200°C. Box 10 could include more than one filter.

[0030] According to another embodiment, the filter 26 is disposed on the box body 12 at the fluid connections 20. According to one embodiment, the filter 26 is disposed on the box body 12 opposite the fluid connections 20. This latter configuration can limit the risk of wetting the filter 26.

[0031] The two fluid inlet connections 22 are ports having an internal tubular extension 40 extending into the interior 14 of the box body 12. Each tubular extension 40 has an end that is vertically above the working surface 32, or if the box has an insert 30, vertically above the culture area 34 of the insert 30. The internal tubular extensions 40 are dimensioned to be long enough to supply the culture medium, but at the same time not so long as to hinder the removal of the insert 30. According to one embodiment, one end 42 of the fluid inlet connections 22 in the interior 14 of the box body 12 is beveled. The beveled shape allows flow towards the culture area 34 / working surface 32. The bevel is for example oriented towards the upper opening 16. The bevel can be approximately 45 degrees.The internal tubular extensions 40 can be sealed to the box body 12 by welding and the bevels obtained using an aseptic disconnecting machine.

[0032] The fluidic connections 20 also each have an external tubular extension 41 extending from the outside of the box body 12 so as to connect to T-tubes containing culture or drainage fluids (shown in dotted lines). The external tubular extensions 41 are adapted to connect to the T-tubes, for example, by a male-female system with a hose clamp that ensures a seal, a mechanical system, by ultrasonic welding, or by using biocompatible glue.

[0033] The inlet fluid connections 22 and / or the outlet fluid connection 24 may be septum-shaped, allowing them to be pierced by a needle while maintaining a tight seal. The inlet fluid connections 22 and / or the outlet fluid connection 24 may be resistant to dimethyl sulfoxide (DMSO) or any other preservation medium injected into the plate at the end of the cell culture process.

[0034] The box 10 could also have external fixings, for example a slot in the outside of the box body 12, to allow the box to be temporarily fixed to a support for feeding the box with culture media. The support can thus hold a plurality of boxes 10 to ensure parallel production of TC cell tissue.

[0035] As mentioned previously, the removable insert 30 includes the culture area 34 adapted to receive and culture the TC cellular tissue and the gripping area 35 allowing the insert 30 to be manipulated, in particular to put it in and / or remove it from the box body 12.

[0036] The culture zone 34 may include a top opening 44 adapted to receive the TC cell culture in order to feed the cells from above, particularly through the inlet fluid connections 22. The culture zone 34 includes a porous base 46 opposite the top opening 16 when the insert 30 is placed in the box body 12 on the working surface 32. The porous base 46 of the insert 30 receives the TC cell culture. It is porous to allow the fluid culture medium to saturate the top and bottom of the TC cell culture. The porous base 46 may be made of polyethylene or polycarbonate. The porous base 46 may be transparent. The porous base 46 may be flexible or rigid. It may have undergone plasma treatment. By porous, it is understood that the base allows the drainage of liquids, either through an open cell network or one or more drainage holes.The porous base 46 may or may not be detachable from the rest of the insert 30. According to one embodiment, the pore size of the porous base is 0.2 to 4 µm, preferably 0.2 to 0.6 µm. According to one embodiment, the pore density of the porous base is 1 x 10⁶ to 5 x 10⁶, preferably 1 x 10⁶ to 3 x 10⁶ pores per cm².

[0037] The growing zone 34 of the insert 30 is generally rounded in shape, for example ring-shaped. The growing zone 34 of the insert 30 may have a thickness E that is less than or equal to the thickness of the working surface 38 of the box body. 12 adapted to receive the culture zone 34 of the insert 30. This can allow good fluid flow. According to one embodiment, the culture zone 34 of the insert 30 has a shape corresponding to a shape of the working surface 32 of the box body 12 adapted to receive the culture zone 34 of the insert 30 to prevent the insert 30 from moving relative to the box body 12 during transport.

[0038] The gripping area 34 is, in one embodiment and as illustrated in the figures, a handle. However, the gripping area 34 could have any shape that allows manipulation of the insert 30. In the embodiment illustrated in the figures, the gripping area 34 is positioned opposite the inclined portion 38. The gripping area 34 may be inclined relative to the horizontal plane H towards the culture surface 32 to allow the flow of condensed fluids, for example. The inclination of the gripping area 34 relative to the horizontal plane H may be substantially the same as the inclination of the inclined portion 38.

[0039] The insert 30 may further include one (or more) connection(s) 47 adapted to cooperate removably with the interior 14 of the box body 12. The connections 47 may prevent the insert 30 from moving relative to the box body 12 during transport. In one embodiment, and as illustrated in the figures, the connections 47 are tabs adapted to be removably inserted into a recess 48 in the interior 14 of the box body 12. The recess 48 may be an open recess to allow the insert 30 to be lifted out.

[0040] The insert 30 may be made of polypropylene, fluorinated ethylene propylene, or polycarbonate. The insert 30 may be resistant to dimethyl sulfoxide (DMSO).

[0041] The box 10 is usable as follows. Initially, the box 10 is supplied with the sealed closure means 18, and the insert 30 inside 14 of the box 10, if the box 10 has such an insert. The insert 30 can contain cells in the working area 36 (i.e., on the working surface 32 if the box does not have an insert, or on the porous bottom 46 if the box has an insert) which will be used to culture the TC cell tissue. The box 10 is then connected via the fluid connections 20 to culture media through the tubes T. There may be one or more culture media which are conveyed into the working area 36 via the inlet fluid connections 22, sequentially, with (or without) rest periods in between. The fluidic outlet connection 24 allows drainage of the working area 36 between the different culture medium connections. The TC cell tissue culture is therefore carried out in a closed environment with minimal risk of contamination.When the TC cell tissue is created, the box 10 can be cryogenically frozen and transported. When the operator decides to use the TC cell tissue, they can retrieve it in several ways. They can open the closure means 18 and retrieve the TC cell tissue. If the box 10 has the insert, they can then remove the insert through the upper opening 16, which is thus freed from the closure means 18.

[0042] According to one embodiment, and as illustrated in [Fig. 6], the porous base 46 of the insert 30 can be detached, if necessary via a tab, so as to extract the TC cellular tissue from the insert 30 without damaging it. Initially, the porous base 46 with the TC cellular tissue on it can be separated from the insert 30, and then the porous base 46 removed from the TC cellular tissue so as to be able to apply the TC cellular tissue alone, for example, to the skin of patient S.

[0043] According to another embodiment and as illustrated in [Fig.7], the porous base 46 is dissociated from the insert 30 and the cellular tissue TC remains for a time attached to the insert 30. The insert 30 is then applied to the skin S of the patient in order to place the cellular tissue TC on the skin S of the patient without manipulating it directly.

[0044] On the other hand, the box 10 could have a removable base for retrieving the TC cell tissue, particularly when the box 10 does not have an insert. Referring to [Fig. 8], the working surface 32 of the box body 12 is removable and preferably flexible. When the TC cell tissue is formed and ready for use, the working surface 32 is removed from the box body 12, along with the TC cell tissue. The TC cell tissue is then removed from the working surface 32 and deposited onto the patient's skin S.

Claims

Demands

1. Cell tissue culture and transport (CT) box (10) comprising: - a box body (12) having an accessible interior (14) and a top opening (16); - a filter (26) disposed in the box body (12) and communicating the interior (14) of the box body with an exterior (28) of the box body (12), the filter (26) being configured to allow gas exchange between the interior (14) and the exterior (28) of the box body (12), while preventing contamination of the cell tissue culture; - a closure means (18) closing the top opening (16) of the box body (12), the closure means (18) being fluid and gas tight; and - at least one aseptic fluid connection (20) through the box body (12), said at least one aseptic fluid connection (20) including two inlet fluid connections (22) for fluids and one outlet fluid connection (24) for fluids.

2. Box (10) according to claim 1, wherein the closing means (18) is single-use such that once detached from the upper opening (16), the interior (14) of the box body (12) is accessible through the upper opening (16).

3. Box (10) according to claim 1 or 2, wherein the closure means (18) is transparent.

4. Box (10) according to any one of the preceding claims, further comprising a removable insert (30) adapted to be disposed in the interior (14) of the box body (12), the insert (30) being adapted to receive a cell tissue culture (CT).

5. Box (10) according to the preceding claim, wherein the insert (30) is removable from the box body (12) through the upper opening (16).

6. Box (10) according to claim 4 or 5, wherein the insert (30) has a culture zone (34) and a gripping zone (35) connected to the culture zone (34), the gripping zone (35) being adapted to manipulate the insert (30) relative to the box body (12).

7. Box (10) according to the preceding claim, wherein the culture area (34) includes a top opening (44) adapted to accommodate the culture of cell tissue (CT).

8. Box (10) according to claim 6 or 7, wherein the growing area (34) includes a porous base (46).

9. Box (10) according to the preceding claim, in which the porous bottom (46) is removable from the rest of the insert (30).

10. Box (10) according to any one of claims 6 to 8, wherein the insert (30) includes at least one connection (47) adapted to cooperate removably with the interior (14) of the box body (12).

11. Box (10) according to any one of the preceding claims, wherein the filter (26) is disposed in an inclined part (38) of the interior (14) of the box body (12).

12. Box (10) according to any one of the preceding claims, wherein the interior (14) of the box body (12) includes a working area (36) adapted to receive cell tissue culture (CT) and fluid culture media enabling the cell tissue (CT) to grow.