Closed device for the culture and transport of cell therapy products

Abstract

The present invention relates to a closed device (1) for culturing and transporting cell therapy products, comprising a container (2) with a base (20) surmounted by peripheral walls (21), delimiting an internal volume with an opening (22) in its upper part; a closure means (3) attached as a removable fastener covering said opening (22); characterized in that at least one of the peripheral walls (21) comprises at least two through channels (4) opening into said internal volume. The invention also relates to a kit (7) equipped with such a culture device (1) and a harvesting organ (8). Figure for the abstract: Fig. 1

Description

Title of the invention: Closed device for the culture and transport of cell therapy products Technical field of the invention

[0001] The present invention falls within the medical field of tissue culture. Tissue culture is performed in vitro and consists of growing tissues or cells in a sterile artificial medium. Tissue culture generally involves a nutrient culture medium, in liquid, semi-solid, or solid form.

[0002] The invention will find preferential application in the culture of cell therapy products of the skin substitute type, such as the epidermis, composed of human pluripotent stem cells, more specifically based on keratinocytes, for example keratinocytes derived from pluripotent stem cells or based on melanocytes, or fibroblasts, or even endothelial cells. The invention further aims at a therapeutic application by grafting animal skin but preferably human, for example in the treatment of skin ulceration.

[0003] For this purpose, the invention relates to a device for culturing and transporting cell therapy products, hereinafter referred to as the "culture device". State of the art

[0004] Currently, cell culture involves a container with a base and peripheral walls, forming an internal volume with an opening at the top, closed by a lid. This internal volume allows the culture medium to be deposited through the opening, followed by the cells to be cultured. Such a container is generally in the form of a Petri dish, cylindrical in shape, or more rarely rectangular parallelepiped in shape. One disadvantage of a Petri dish lies in the closure of its lid, which is not airtight; this allows the gas exchange necessary for the proper culture of cells, but does not in any way ensure maintenance under a sterile atmosphere.

[0005] An alternative solution consists of a container in the form of a culture flask. Such a flask is generally flask-shaped, with a top opening hermetically sealed by a lid. This flask also includes, on one side wall, an upward-sloping neck with a spout fitted with a cap, which can be screwed from a hermetically sealed position to a venting position, and vice versa. Such a cap may also include a filter membrane, generally hydrophobic, protecting the inside of the flask from contaminants present in the outside atmosphere. Less commonly, the cover may be replaced by a peelable film, sealing the opening airtight. Such a film is applied by sealing, particularly heat sealing, to the edges or rims of the surrounding walls around the opening.

[0006] Furthermore, these containers are made of plastic material, the characteristics of which allow sterilization, but also the good development of the cells to be cultured within such containers.

[0007] These various known containers present numerous drawbacks, due to their formats and dimensions, and whether or not they are airtight, making their transport either difficult or requiring meticulous monitoring, or impossible. In fact, non-airtight containers require maintenance under a controlled atmosphere. Furthermore, they require transport with extreme precautions, making them impractical. These containers also impose restrictions on introducing multiple solutions one after the other without contamination. Description of the invention

[0008] The invention aims to overcome the drawbacks of the prior art by providing a closed device for culturing and transporting cell therapy products. Such a culture device has features and various elements that internally ensure the development of the cell culture in a protected medium, hermetically sealed by a lid or hermetically covered by a peelable film, while allowing the introduction and extraction of various solutions required for said culture, in a controlled manner and without risk of contamination. In addition, the cultivation system allows for easy transport of crops, without restrictive precautions.

[0009] To achieve this, the cultivation device includes - a container with a base surmounted by peripheral walls, delimiting an internal volume with an opening at the top; - a closing mechanism, attached as a removable fastener covering said opening;

[0010] characterized in that - at least one of the peripheral walls includes at least two through channels opening into said internal volume.

[0011] According to additional, non-limiting features, such a cultivation device

[0012] may include - two primary channels, formed through a first of the peripheral walls; - three secondary channels, formed through a second of the peripheral walls. According to one embodiment, said primary and secondary channels are equipped with plugs dedicated to: i) a filtration allowing gas exchange; j) an inlet allowing the introduction of a culture medium; jj) an entry allowing the introduction of a hydrogel matrix of exclusively biological origin; jjj) an entry allowing the introduction of cells; k) an outlet allowing the evacuation of said culture medium.

[0013] According to one embodiment, said device comprises - a cutting tool sized and shaped for insertion within said internal volume until it comes into contact with said bottom; - the cutting tool being provided on the underside with at least one cutting module with beveled lower edges forming cutting blades.

[0014] According to one embodiment, said device includes a pusher, attached to the upper face of said cutting tool.

[0015] According to one embodiment, said at least one cutting module has an overall rectangular parallelepiped shape, with a truncated corner.

[0016] According to one embodiment, said cutting tool comprises six cutting modules distributed in two rows and three columns.

[0017] According to one embodiment, said closure means is airtight and comprises a cover, provided with lateral tabs for snap-fitting attachment with said peripheral walls.

[0018] According to one embodiment, said closure means comprises a peelable lid attached by heat-sealing to the upper edge of said peripheral walls.

[0019] The invention further relates to a cell therapy product culture and transport kit comprising - a cultivation device according to the invention, characterized in that it comprises - a sampling device shaped like a shovel, sized to complement said cutting tool, - said shovel comprising a handle connected to a flat end, equipped with - said flat end of said shovel includes a rough surface. Presentation of the drawings

[0020] Other features and advantages of the invention will become apparent from the description detailed, non-limiting embodiments of the invention, with reference to the attached figures, in which:

[0021] [Fig. 1] schematically represents a perspective view of an embodiment of a culture kit, showing in particular said culture device with a pusher and two formats of cutting tools, as well as the lid and the sampling organ in the form of a shovel;

[0022] [Fig.2] schematically represents a perspective view of an embodiment of said culture device, closed by its lid and with five primary and secondary channels connected to different plugs;

[0023] [Fig.3] schematically represents a view similar to 1 a [Fig.2], showing the culture device hermetically sealed by a capped film;

[0024] [Fig.4] schematically represents a perspective view of several stacked cultivation devices;

[0025] [Fig.5] schematically represents a perspective view of the underside of a format of the cutting tool with six cutting modules, showing in particular the beveled edges of each of said six cutting modules;

[0026] [Fig.6] schematically represents a perspective view of an embodiment of the pusher, showing in particular an impact face provided with six cylindrical lugs;

[0027] [Fig.7] schematically represents a view similar to [Fig.6], with said cultivation device, within the housing of which is introduced a cutting tool with six cutting modules. Detailed description

[0028] The present invention falls within the medical field of tissue culture and relates to a device 1 for culturing and transporting cell therapy product, hereinafter referred to as "device".

[0029] Such a device 1 is intended to be closed, namely that it has an internal volume accessible during certain operations, such as the taking of a cultured sample, but which can be closed again, in order to define a closed environment, separate from the outside atmosphere.

[0030] Further on, the device 1 comprises a container 2 with a base 20 surmounted by peripheral walls 21, delimiting an internal volume with an opening 22 in the upper part.

[0031] Such a container 2 can have any shape, preferably rectangular parallelepiped, in particular with rounded corners. In addition, container 2 has a generally flat base 20, in order to allow for the development of tissue culture.

[0032] According to a preferred embodiment, in order to facilitate storage and transport, several devices 1 can be stacked. To achieve this, the said peripheral walls 21 are shaped in an inverted U with a hollow. In other words, the said peripheral walls 21 of a first device 1 have an internal space, open at the bottom, allowing the insertion and fitting of an upper part of the peripheral walls 21 of another device 1 thus fitted together. In particular, said walls 21 include an upper edge forming an overhang 210 parallel or substantially parallel to said bottom 20. In short, the upper edge of the peripheral walls 21 is flat. Furthermore, projecting from said overhang 210, said peripheral walls 21 include an offset 211 shaped complementary to said recess, forming a stacking lug for several cultivation devices 1. In particular, said offset 211 is positioned recessed from the outer face of the peripheral walls 21, with a portion of the overhang 210 forming a bearing surface for the lower edge of the peripheral walls 21 of a superimposed device 1. Furthermore, the peripheral walls 21 may project beyond the bottom 20, extending below, which allows several devices 1, whose container 2 is closed, to be nested one on top of the other, in particular by a lid 30 Such an offset 211 extends over all or part of the upper edge of the peripheral walls 21, preferably at least along two adjacent walls 21. The said offset 211 may be continuous or interrupted, preferably continuous at the levels of at least two corners of said device 1. According to a preferred embodiment, as seen in [Fig.3], said offset 211 extends continuously along two adjacent peripheral walls 21, as well as over part of the length of the other two walls 21, with rounded corners. In addition, the offset 211 can serve as a keying device, with a view to vertical alignment along the same orientation of at least two devices 1 thus nested. Figure 4 shows three devices 1 superimposed and oriented in the same direction, in particular three stacked empty containers 2.

[0033] As mentioned previously, in order to determine an internal closed space suitable for tissue culture, the device 1 is provided to be recloseable and includes for this purpose a closure means 3, brought in as a removable fixing covering said opening 22.

[0034] According to an advantageous embodiment, said closure means 3 is hermetically sealed and comprises a cover 30, provided with lateral tabs 31 for snap-fit ​​attachment to said peripheral walls 21. For this purpose, the peripheral walls 21 The spherical components may include any type of complementary element, shaped to cooperate by snapping with said tabs 31. Therefore, in the snapped position of the lateral tabs 31, the cover 30 provides a complete seal over the opening 22, against both air and any fluid. Furthermore, the underside of said cover 30 may include seals ensuring a seal against the surface of the walls of the container 2 and by compression upon snapping. As can be seen in [Fig.2], the tabs 31 can be snapped into place at the outer edges of the peripheral walls 21. According to a preferred embodiment, as shown in Figures 1 and 2, the cover 30 comprises two tabs 31, formed at two adjacent edges of said cover 30. The cover 30 is then positioned at the diametrically opposite corner, interlocking internally with the offset 211 projecting from the upper edges of said peripheral walls 21. The cover 30 then, on the one hand, interlocks with said offset 211 at two edges and, on the other hand, clips into place by means of its tabs 31, which engage with the two opposite edges, ensuring its positioning and retention in the closed position of said opening 22. In addition, said cover 30 may include on its underside means ensuring sealing in the closed position, such as seals provided on the perimeter of said underside of said cover 30.

[0035] According to a preferred embodiment, said closure means 3 comprises a peelable lid 32 attached hermetically by heat-sealing to the upper edge of said peripheral walls 21. Such a lid 32 must ensure a seal against contaminants and fluids, from both the outside and the inside, while allowing the gas exchange necessary for the proper development of the culture. According to a preferred embodiment, the heat-sealing of said lid 32 is carried out on the overhang 211 formed on the upper edge of the peripheral walls 21.

[0036] Furthermore, said lid 32 can be made of any type of material, preferably a composite or plastic material, such as a polyethylene plastic. This material can be transparent or translucent, allowing light to pass through and enabling observation inside the volume of the device 1, for example, to monitor the development of the tissue culture. Such a material can also include a filtering coating, limiting or blocking certain wavelengths of light radiation, such as certain ultraviolet rays, which are harmful to the development of certain cells. Figure 3 shows an example of a lid 32 in the form of a transparent plastic film, heat-sealed onto said overhang 211. This embodiment shows in particular a part of the overhang 210 provided internally in relation to said overhang 211, forming a flat surface around the internal volume of the container 2.

[0037] According to a combined embodiment, the cover 30 can be attached over the lid 32.

[0038] Advantageously, the invention allows the introduction and extraction of different solutions required for said culture, in a controlled manner and without risk of contamination, in particular by keeping the internal volume hermetically sealed. To achieve this, device 1 provides at least one of the peripheral walls 21 comprising at least two channels 4 passing through and opening into said internal volume. The said at least two channels 4 may be located at the level of the same peripheral wall 21, or at the level of two different peripheral walls 21, preferably adjacent. Each channel 4 includes a portion that protrudes externally from the corresponding peripheral wall 21.

[0039] According to a preferred embodiment, the culture device 1 comprises two primary channels 40,41, formed through a first of the peripheral walls 21, as well as three secondary channels 42,43,44, formed through a second of the peripheral walls 21. Each of the primary channels 40, 41 and the secondary channels 42, 43, 44 are then dedicated to the introduction or extraction of a solution, as well as to gas exchange.

[0040] According to one embodiment, the primary channels 40, 41 and the secondary channels 42, 43, 44 are equipped with plugs 400. Such plugs 400 are attached with a removable fixing, in particular by fitting them into the protruding portion of the channels 4, thus forming a tip. Such a tip is preferably of the "LUER" or "LUER LOCK" type, an international standard in the medical field, corresponding to a 6% taper, possibly with tensile-resistant serrations, and a screw system provided on the plugs, allowing connection with another medical device or organ. Further on, the aforementioned 400 caps are dedicated to: i) a filtration allowing gas exchange; j) an inlet allowing the introduction of a culture medium; jj) an entry allowing the introduction of a hydrogel matrix of exclusively biological origin; jjj) an entry allowing the introduction of cells; k) an outlet allowing the evacuation of said culture medium. A device 1 equipped with five 400 plugs is visible in [Fig.2]. According to one embodiment, said channels 4 can be connected to an automated or semi-automated culture system. Said plugs 400 can therefore include fittings for tubes or tubing, or even a septum provided with a self-sealing membrane, generally made of silicone material (the material and thickness of this membrane allowing it to be pierced by a needle, and especially to retain its airtight character once the needle is removed). In particular, i) the filtration allowing exchange can be equipped with a plug 400 provided with an elbow on which is mounted a filter, notably at 0.22 µm, filtering particles and fluids, but allowing gas exchange with the atmosphere; j) the inlet of the culture medium may be fitted with a plug 400 provided with a safety non-return valve, to prevent the medium from escaping to the outside once introduced through this channel 41, particularly during transport and handling; jj) the matrix inlet, such as hydrogel), is used only once and includes a 400 cap which is intended to be sealed; jjj) the cell inlet, during seeding, is used only once and includes a 400 plug which is intended to be sealed; (k) The outlet of the "consumed" medium includes a stopper 400 equipped with a safety non-return valve to prevent the medium from flowing back into the device during extraction.

[0041] Thus, during the use of the device 1 for tissue culture, first, after sterilization and sealing of the container 2, in particular with the film 32, the matrix is ​​injected using a syringe, in particular a standardized "LUER LOCK" type syringe, into the secondary channel 42. The air inside is purged by gently pressing on the film 32 before sealing said secondary channel 42 with a stopper 400. The matrix is ​​processed for a period of two (2) hours at a temperature of 37.5°C (degrees Celsius), in particular in an incubator. Several devices 1 can then be covered with their lids 31 and stacked for simultaneous incubation. This incubation allows in particular coagulation and adhesion to the bottom 20 of container 2. Next, a cell suspension is injected using a Luer Lock syringe through the secondary channel 43. Air is purged by gently pressing on the film 32 before the channel 43 is closed with a stopper 400. Device 1 containing the matrix and cell suspension is then treated for 24 h at a temperature of 37.5°C, in particular within an incubator), to allow the adhesion of cells to the matrix. Then, periodically, every other day until 7 days are reached, the "consumed" medium contained in device 1 is evacuated using a "LUER LOCK" type syringe via the secondary outlet channel 44. Fresh medium is injected using from a "LUER LOCK" type syringe via the primary inlet channel 41. Device 1 is brought to a temperature of 37.5°C, in particular within an incubator, until the next change of medium. At the end of the procedure, the used medium from device 1 is evacuated using a Luer-lock syringe via the secondary outlet channel 44. Fresh medium is injected using a Luer-lock syringe via the primary inlet channel 41. The filter is removed, and channel 40 is sealed with a suitable stopper 400, similar to channel 4L. The lid 31 is then clipped onto the device 1, hermetically sealing the assembly, protecting the lid 32: the device 1 is then ready to be transported.

[0042] According to one embodiment, the device 1 includes a cutting tool 5 dimensioned and shaped for its insertion within said internal volume until it comes into contact with said bottom 20. In other words, the tool 5 has smaller dimensions, in length and width, compared to the internal dimensions of the container 2, to allow its insertion and extraction within said internal volume, until it comes into contact with the surface of said bottom 20. In addition, the cutting tool 5 is provided on its underside with at least one cutting module 50 with lower beveled edges 51 forming cutting blades. According to one embodiment, as seen in Figures 5 and 7, said cutting tool 5 comprises six cutting modules 50 distributed in two rows and three columns. Thus, it is possible to divide the tissue culture into several portions or "grafts", in particular into as many grafts as there are 50 modules.

[0043] It should be noted that the bevel of the lower edges 51 of each cutting module 50 forms an inclination with the corresponding vertical wall. The inclination of this bevel can be oriented inwards or outwards. In addition, the thickness of each edge 51 is minimal, forming a blade edge capable of penetrating and cutting through the tissue culture.

[0044] Further, according to one embodiment, each cutting module 50 has a generally rectangular parallelepiped shape. Preferably, each cutting module 50 has a rectangular parallelepiped shape, with a truncated corner, used to cut a notch in each graft and determining a visually recognizable orientation of said graft, in particular to ensure that the keratinocytes are positioned superiorly during manipulation of said graft, or to notice if it has turned over and to be able to put it back in the correct orientation, of the cutting tool 5 within the device 1. In particular, the truncated corner of a module 50 located at the periphery, near a corner of the internal volume of the container 2, can then serve as a keying feature.

[0045] According to one embodiment, the device 1 comprises a pusher 6, attached to the upper face of said cutting tool 5. Such a pusher 6 serves as a surface to ensure, on the one hand, the gripping of said cutting tool 5 during its insertion and extraction within the container 2 and, on the other hand, a bearing surface in a downward vertical direction in order to apply sufficient pressure to ensure the cutting of the tissue culture. In particular, according to a preferred embodiment, the cutting can be carried out by jerking, by hammering the upper face of said pusher 6, for example with a surgical mallet. To achieve this, said pusher 6 includes on its upper face a knocker, in the form of at least one impact contact face provided with solid lugs 61. According to one embodiment, as seen in figures 1 and 6, said lugs 61 have a cylindrical shape. The knocker may include one or more lugs 61, preferably six lugs 61 arranged in two rows and three columns, corresponding to the embodiment with six cutting modules 50. These modules are then vertically aligned with said six lugs 61 when said knocker is used with said cutting tool 5, improving the pressure applied for cutting each of said modules 50. Accordingly, according to one embodiment, said pusher 6 can be removably attached to the cutting tool 5, in particular by additional fitting. According to the embodiment shown in [Fig.1], the pusher 6 has a shoe 60, projecting on the lower face and dimensioned complementaryly, except for the clearance, to be inserted by sliding fit into a recess 52 which is provided by the cutting tool 5 on the upper face. According to a preferred embodiment, said shoe 60 is provided with snap-on means for clipping with said impression 52, ensuring the locking of these two elements together. Thus, the pusher 6 mounted with the cutting tool 5 form two integral elements, whose lugs 6 ensure in particular resistance to impacts during hammering, as well as the integral transmission of the force thus applied, in order to ensure the cutting of the crop by the modules 50.

[0046] According to one embodiment, as seen in [Fig.6], the lugs 61 can be reinforced by connections 62, improving the rigidity of said pusher 6. In addition, similar reinforcements can be provided on the inner face of said pusher 6. It should be noted that the lugs 61 and the links 62 can be made in an integral way during the molding of the plastic part of said pusher 6.

[0047] According to an advantageous embodiment, the device 1 is made of material plastic. In other words, the various elements of said device are made of plastic material, namely the container 2, the lid 30, or the cutting tool 5, or the pusher 6. Preferably, device 1 is made of polypropylene (PP), in particular "HP671T" polypropylene. According to one embodiment, said cutting tool 5 and / or pusher 6 are made of composite or plastic material, ensuring increased resistance to impacts and providing a sufficiently strong and sharp blade edge for each cutting module 50.

[0048] According to one embodiment, all or part of the device 1 is made of PP that has undergone plasma treatment. Such plasma treatment makes it possible to modify the Arithmetic Mean Roughness (Ra) of the surface of the device 1, in particular the internal walls and especially the bottom 20 of the container 2, increasing its hydrophilic properties, without generating particles or leachable substances. Plasma-treated PP plastic thus provides a surface suitable for coagulation and fibrin adhesion. Furthermore, the PP plastic material ensures that the characteristics of device 1 are maintained during sterilization, particularly during sterilization by irradiation. It should be noted that the PP plastic material also allows the sealing of the lid 32. In addition, the PP plastic material provides resistance to impacts during cutting.

[0049] The invention also relates to a cell therapy product culture and transport kit 7, comprising a culture device 1 according to the invention, as previously described. An example of kit 7, with device 1 and its various parts, can be seen in particular on [Fig.1]. Advantageously, such a kit 7 includes a sampling organ 8 shaped in the form of a shovel, dimensioned complementaryly to said cutting tool 5. According to one embodiment, said shovel comprises a handle 80 connected to a flat end 81. In particular, said flat end 81 has dimensions substantially equal to those of a module 50 of the cutting tool 5, preferably slightly larger dimensions, in order to ensure the removal of a previously cut graft, with sufficient clearance. According to a preferred embodiment, said handle 80 is grooved, improving grip and preventing it from slipping in the hands.

[0050] According to an advantageous embodiment, said flat end 81 of the shovel includes a rough surface, improving the adhesion of the graft taken and preventing it from slipping when it is taken. According to a preferred embodiment, said flat end 81 includes an inclined distal end, allowing the bottom 20 to be scraped and a graft to be recovered. The flat end 81 also includes two raised longitudinal edges, preventing the graft from being taken from sliding laterally during the taking or moving of said shovel. An example of such a shovel-shaped organ 8 for harvesting is notably visible in [Fig.1].

[0051] According to one embodiment, said sampling organ is made entirely of plastic material, preferably polypropylene (PP).

[0052] Further, according to one embodiment, the device 1 has dimensions allowing the culture and transport of a tissue culture with a surface area of ​​approximately 60 square centimeters (cm2), which can be divided during cutting into grafts of approximately 10 cm2, directly within the device 1 without extracting said culture.

Claims

Demands

1. A closed device (1) for culturing and transporting a skin substitute cell therapy product, comprising: - a container (2) having a rectangular parallelepiped shape with a generally flat base (20) surmounted by peripheral walls (21), delimiting an internal volume with an opening (22) in its upper part; - a closure means (3), attached as a removable fastener covering said opening (22); - at least one of the peripheral walls (21) comprising at least two through channels (4) opening into said internal volume; characterized in that it comprises: - a cutting tool (5) dimensioned and shaped for its insertion into said internal volume until it comes into contact with said base (20); - the cutting tool (5) being provided on its lower face with at least one cutting module (50) with beveled lower edges (51) forming cutting blades;- said at least one cutting module (50) having an overall rectangular parallelepiped shape, with a truncated corner.;

2. Culture device according to the preceding claim, characterized in that it comprises - two primary channels (40,41), formed through a first of the peripheral walls (21); - three secondary channels (42,43,44), formed through a second of the peripheral walls (21).

3. Culture device (1) according to the preceding claim, characterized in that - said primary and secondary channels (40,41,42,43,44) are equipped with plugs dedicated to: i) a filtration allowing gas exchange; j) an inlet allowing the introduction of a culture medium; jj) an inlet allowing the introduction of a hydrogel matrix of exclusively biological origin; jjj) an inlet allowing the introduction of cells; k) an outlet allowing the evacuation of said culture medium.

4. A cultivation device (1) according to any one of the preceding claims, characterized in that it comprises - a pusher (6), attached to the upper face of said cutting tool (5).

5. Cultivation device (1) according to any one of the preceding claims, characterized in that said cutting tool (5) comprises - six cutting modules (50) distributed in two rows and three columns.

6. A culture device (1) according to any one of the preceding claims, characterized in that - said closure means (3) is airtight and comprises a lid (30), provided with lateral tabs (31) for snap-fit ​​attachment with said peripheral walls (21).

7. Culture device (1) according to any one of the preceding claims, characterized in that - said closure means (3) comprises a peelable lid (32) attached hermetically by heat-sealing to the upper edge of said peripheral walls (21).

8. Kit (7) for culturing and transporting cell therapy product, comprising - a culture device (1) according to any one of claims 1 to 7, characterized in that said kit (7) comprises - a sampling organ (8) shaped in the form of a shovel, dimensioned complementary to said cutting tool - said shovel comprising a handle (80) connected to a flat end (81), provided with a rough surface.

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