Valve system for cell culture automation.

Abstract

The present invention relates to a device (1) for controlling the passage of a liquid in a series of flexible tubes, comprising: - a set of clamps (20), each being capable of clamping a flexible tube, and comprising a first jaw (21) arranged on a lever (30) and a second jaw (22) forming a jaw with said first jaw (21), each lever (30) being pivotally mounted between an open position in which said clamp (20) is open and a closed position in which said clamp (20) is closed, and - a set of wheels (40) mounted integrally in rotation about the same axis A, each of said wheels (40) forming a cam and being associated with one of said levers (30), characterized in that the rotation of each wheel (40) allows the passage of said lever (30) from its closed position to its open position and vice versa.

Description

Title of the invention: Valve system for cell culture automation. Technical field of the invention

[0001] The present invention belongs to the field of scientific apparatus. More particularly, the present invention belongs to the field of fluid circulation control. The device according to the invention is more particularly intended for managing the distribution of culture medium to cell culture dishes. State of the art

[0002] Cell culture is an essential element of biological research. It is used in particular for the study of cellular behavior and the production of recombinant proteins or viruses. For this purpose, adherent or suspended cells are cultivated in a liquid medium inside gas- and temperature-regulated enclosures.

[0003] Outside of industrial production devices, cell culture requires numerous human manipulations. The cells are cultivated in dishes placed in a dedicated incubator. To feed the cells and remove the waste produced, an operator must regularly renew the culture medium by removing used medium and reintroducing fresh medium. In addition, the cell density must be controlled. Too low or too high a density prevents their homogeneous growth. Consequently, the operator must regularly remove cells in excess quantity to discard them or distribute them in other culture dishes. All these manipulations consume time (including weekends and holidays) and materials (e.g. pipettes). In addition, the regular opening and closing of culture dishes and bottles containing the culture medium are significant sources of bacterial contamination.

[0004] There is therefore a significant need to eliminate human intervention as much as possible in the process of in vitro cell culture.

[0005] Automated devices are already commercially available. However, they cost several hundred thousand euros to install and require many dedicated consumables. These devices are suitable for industrial production but not for the scale of the research and development laboratory.

[0006] Ideally, a cell culture machine should be able to adapt to the equipment already existing inside the laboratory. Thus, it must be compact and lightweight to fit into the incubators alongside the culture dishes. Alternatively, the machine must be able to be placed next to the incubator. It is advantageously modular to be able to adapt to different configurations.

[0007] Among the central modules of a cell culture machine, we can cite the pump(s) which will supply the culture dishes with medium as well as the valves which will control the passage of liquid towards the dishes and / or towards the emptying or cell collection containers.

[0008] To facilitate cleaning and avoid contamination, the valves must not be in direct contact with the culture medium. This latter constraint excludes the use of rotary valves.

[0009] Furthermore, the valves must be light, compact, easy to manufacture and low energy consumption. The use of a pinch solenoid valve is therefore not very feasible in this field.

[0010] There therefore remains a need for a valve system, adapted to a cell culture machine meeting the constraints described above. Summary of the invention

[0011] Thus the present invention relates to a device for controlling the passage of a liquid in a series of flexible tubes, comprising: - a set of clamps, each being capable of clamping a flexible tube, and comprising a first jaw disposed on a lever and a second jaw forming a jaw with said first jaw, each lever being pivotally mounted between an open position in which said clamp is open and a closed position in which said clamp is closed, and - a set of wheels mounted integrally in rotation around the same axis A, each of said wheels forming a cam and being associated with one of said levers, characterized in that the rotation of each wheel allows the passage of said lever from its closed position to its open position and vice versa.

[0012] According to a preferred embodiment of the invention, said device comprises a first stop limiting the movement of said lever in the open position and a second stop limiting the movement in the closed position.

[0013] In the context of the present invention, the expression "capable of clamping a tube" is intended to designate the fact that said clamp is capable of clamping at least one tube. In a preferred embodiment of the invention, said clamp is capable of clamping a single tube or two tubes.

[0014] According to a preferred embodiment of the invention, each lever is constrained in the closed position via an elastic means.

[0015] According to a preferred embodiment of the invention, the elastic means is a spring.

[0016] According to a preferred embodiment of the invention, said second jaw is fixed.

[0017] According to a preferred embodiment of the invention, said first and second jaws two adjacent clamps are arranged in a staggered pattern.

[0018] According to a preferred embodiment of the invention, each second stop is arranged so that said first and second jaws are not in contact in the closed position.

[0019] According to a preferred embodiment of the invention, said first and second jaws are separated by a space equivalent to that of the thickness of a flexible tube when it is crushed.

[0020] According to a preferred embodiment of the invention, each second stop is arranged such that each lever does not bear on the cam path when said lever is in the closed position.

[0021] According to a preferred embodiment of the invention, each lever comprises an arm which extends perpendicular to the axis of rotation A.

[0022] According to a preferred embodiment of the invention, each lever is pivotally mounted around an axis B parallel to the axis of rotation A.

[0023] According to a preferred embodiment of the invention, each lever comprises an arm from which an L-shaped blade (tab) extends perpendicularly.

[0024] According to a preferred embodiment of the invention, each lever comprises at its distal end a tab intended to allow the clamp to be opened by a user.

[0025] According to a preferred embodiment of the invention, each wheel comprises two parallel planar faces delimiting a cam profile comprising a neutral zone, an ascending zone, a plateau zone and a descending zone.

[0026] According to a preferred embodiment of the invention, said set of wheels is associated with a motor with determination of the angular position (60) capable of driving said set of wheels in rotation around the axis A. Brief description of the figures

[0027] [Fig-1] represents a side view of an embodiment of a device according to the invention.

[0028] [Fig.2] represents a side view of an embodiment of a device according to the invention.

[0029] [Fig.3] represents a side view of an embodiment of a device according to the invention.

[0030] [Fig.4] represents a top view of an embodiment of a device according to the invention. Detailed description of the invention

[0031] The device 1 comprises a set of clamps 20, each being capable of clamping a flexible tube in which culture medium circulates, and comprising a first jaw 21 arranged on a lever 30 and a second jaw 22 forming a jaw with said first jaw 21. Each lever 30 is pivotally mounted between an open position in which said clamp 20 is open, during which the culture medium can flow, and a closed position in which said clamp 20 is closed and the culture medium can no longer circulate.

[0032] The change of position of each of the clamps 20 is controlled by a wheel 40 belonging to a set of wheels 40 mounted integrally in rotation around the same axis A, each of said wheels 40 forming a cam is associated with one of said levers 30.

[0033] The device 1 according to the invention is in particular intended to be arranged between the source of culture medium and the culture dishes and / or the dishes and the collection containers. Alternative configurations are conceivable. In certain embodiments, it is possible to connect the sources of medium to a buffer container in which the source media are mixed. In this latter embodiment, the device 1 according to the invention, arranged between the sources of medium and said buffer container, makes it possible to precisely measure the quantity of each of the source media.

[0034] The embodiment presented below comprises two devices 1 according to the invention, a first arranged between the source of culture medium and the culture dishes and a second arranged between the culture dishes and a container for collecting used media or cells. Other embodiments are possible using two head-to-tail devices 1 arranged between the sources of culture medium and the culture dishes making it possible to direct K sources of culture medium to N dishes. In the same way, two head-to-tail devices 1 can be arranged between the culture dishes and containers for collecting used media or cells, making it possible to direct N culture dishes to J containers for collecting used media or cells.

[0035] The culture medium can be arranged in enclosures each connected to a first tube intended to channel the culture medium from said enclosure.

[0036] The embodiment described below comprises a single source of culture medium. However, it is entirely possible to use several sources of culture medium in order to be able to increase the quantity of available medium or to be able to use several different culture media intended, alone or in a mixture, for a particular cell type.

[0037] Said first tube communicates with a series of flexible tubes intended to supply the culture boxes with culture medium.

[0038] The connection between said first tube and said series of flexible tubes can be made via an intermediate piece comprising an inlet, intended to be connected to said first tube, and a series of outlets each intended to be connected to one of the tubes. flexible from the soft tube series.

[0039] Said intermediate piece may take the form of a piece comprising, between its inlet and its series of outlets, a set of channels connecting said inlet to each of the outlets so that the liquid coming from said inlet is distributed between each of the outlets.

[0040] Said intermediate piece can in particular be obtained by molding or additive printing. It is preferably made of plastic. In the latter case, it is understood that said intermediate piece can be manufactured at a limited cost and thus constitute a replaceable consumable after each cycle of use.

[0041] The circulation of the culture medium from its source to the cell culture dishes can be generated by a pump placed at any point in the circuit.

[0042] In order to evacuate the used culture medium from the culture dishes to a container for collecting the used media or cells, another device 1 according to the invention may be arranged between said container for collecting the used media or cells and the culture dishes. Alternatively, the same device 1 as that implemented between the source of culture medium and the culture dishes may be used for this purpose.

[0043] In a manner similar to that described above for the circulation of the culture medium from its source to the culture dishes, the used medium is taken from each of the dishes via a flexible tube included in a series of flexible tubes. The latter can be connected to an intermediate piece as described above comprising a series of inlets all communicating with the same outlet connected to the container for collecting the used media or cells.

[0044] The circulation of the culture medium from the cell culture dishes to the container for collecting the used media or cells can be generated by a pump placed downstream of said dish. Said pump can be identical or different from that used for the circulation of the culture medium from its source to the cell culture dishes.

[0045] The device 1 according to the invention will make it possible to open or close the passage of liquid in each flexible tube included in the series of flexible tubes. Said closing is caused by the crushing of said flexible tube between two jaws 21, 22 of the same clamp 20.

[0046] The clamps 20, of the clamp set 20, comprise: - a first jaw 21 arranged on a lever 30 and, - a second bit 22 forming a jaw with said first bit 21.

[0047] The two jaws 21, 22 can take various shapes. Preferably, each of the two jaws 21, 22 has, in straight section, a dome shape whose point extends towards the complementary jaw of the same jaw.

[0048] The lever 30 has a distal end and a proximal end between which extends an arm 31. The proximal end comprises a housing capable of receiving a rod forming a pivot axis B. The latter advantageously has a longitudinal axis perpendicular to the axis of the arm 31.

[0049] The distal end may advantageously comprise a tab 33 intended to allow the clamp to be opened by a user. Said tab 33 allows the user to disengage the clamp 20 in the open position to place one of the flexible tubes of the series of flexible tubes between the jaws 21, 22.

[0050] The first jaw 21 may be arranged on the surface of the arm 31. Alternatively, it may be offset vertically or below said arm 31. In this latter embodiment, the offset of the jaw 21 may be achieved via a blade 32 extending from the arm 31. Advantageously, the arm has an L shape on the base of which the first jaw 21 is placed.

[0051] The lever 30 further comprises a follower part 34 intended to interact with the cam profile. The follower part 34 is advantageously a stud extending from the arm 31 towards the cam profile.

[0052] Preferably, the lever 30 is a single-piece part. Said lever arm 30 can be manufactured by molding or by additive printing.

[0053] The second jaw 22 of the clamp 20 may be fixed or movable. Preferably, said jaw 22 is fixed. By "fixed", we mean the fact that when the jaw moves from the open position to the closed position, the second jaw 22 is stationary, the jaw closing only by bringing the first 21 towards the second jaw 22. In this latter embodiment, the second jaw 22 is advantageously arranged on the path intended to receive the flexible tube.

[0054] The movement of the jaws 21, 22 between said open position and said closed position can be advantageously limited by a first stop 51 limiting the movement of said lever in the open position and a second stop 52 limiting the movement in the closed position. Advantageously, the stops 51, 52 are arranged so as to limit the movement of the lever 30 and are even more advantageously associated with the chassis 70.

[0055] Preferably, the stop 51 is arranged so that the space between the jaws 21, 22 is, when the lever 30 is in contact with said stop 51, greater than the thickness of a flexible tube at rest. The presence of said stop 51 makes it possible to prevent the operator from opening the jaw too far and damaging the elastic means.

[0056] Preferably, the space between the jaws 21, 22 is, in the open position when the open position is caused solely by the wheel 40, between 95 and 99.9% of the thickness of a flexible tube at rest. Thus, in the open position, the flexible tube is held by friction between the two jaws 21, 22 of the jaw and cannot escape from the device 1 according to the invention. By “at rest” is meant the state of said tube, under normal temperature and pressure conditions, in the absence of specific pressure on its surface.

[0057] Preferably, the second stop 52 is arranged so that said first and second jaws 21, 22 are not in contact in the closed position. It is thus understood that the presence of the stop 52 makes it possible to limit the pressure exerted by the two jaws on the flexible tube in the closed position and the risks of wear, or even rupture, of the flexible tube at the contact zone with the stop 21, 22. Even more advantageously, the stop is arranged so that said first and second jaws 21, 22 are separated by a space less than the thickness of a flexible tube when it is crushed and preferably between 50 and 95% of the thickness of a flexible tube when it is crushed. By "thickness of a flexible tube when it is crushed", is meant a thickness substantially equivalent to the thickness of a wall of the flexible tube multiplied by two.Thus, the maximum crushing of the flexible tube allows the creation of a sealed zone preventing the passage of a fluid without damaging the flexible tube.

[0058] By default, each lever 30 is advantageously constrained in the closed position via an elastic means. The latter will apply a force to the lever 30 which is thus maintained in contact with the stop 52. Preferably, a first end of said elastic means is associated with the chassis 70 containing the device 1 according to the invention while the other end is associated with the lever 30. Said elastic means can take any form known to those skilled in the art such as for example leaf springs and coil springs.

[0059] The wheel 40 forming a cam advantageously comprises two parallel flat faces delimiting a cam profile. The latter is preferably broken down into several successive zones. A first zone, called the “neutral” zone, in which the cam profile does not modify the position of the lever 30 held in the closed position under the effect of the elastic means. According to a less preferred embodiment, in the absence of said elastic means, the “neutral” zone can induce the passage and maintenance of the lever 30 in the closed position. The cam profile then comprises a second zone, called the “ascending” zone, in which the cam profile interacts, by direct or indirect pressure, on the lever 30 to make it pass from its closed position to its open position. A third zone, called the “plateau” zone, during which the cam profile interacts, by direct or indirect pressure, on the lever 30 to keep it in the open position.A fourth zone, called the “descending” zone in which the cam profile interacts, by direct or indirect pressure, on the lever 30 to move it from its open position to its closed position. Said ascending zone is arranged upstream of the neutral zone so as to obtain a cycle where these four zones follow one another indefinitely by rotation of . the wheel 40 forming a cam. Said cam profile can be obtained in particular by using an ovoid cam profile with a flat top. The latter corresponding to said plateau zone.

[0060] Advantageously, the profile of the descending and ascending zones are adjusted to offer the least resistance to the levers and thus reduce the torque of the motor used for the rotation of the wheels 40 forming cams. This allows the device 1 according to the invention to use motors of smaller size, weight and consumption.

[0061] The cam profile and its different zones are adapted to the position of the stops 51, 52. A person skilled in the art is capable of determining the most suitable cam profile which will allow the lever 30 to move from its closed position to its open position without having to force the stops 51, 52.

[0062] The plate is deliberately flat, providing a locally stable position for the follower part 34 of the lever 30, making it possible to cut off the power supply to the motor most of the time, without risk of the wheel slipping.

[0063] Preferably, the stop 52, the cam profile and / or the follower part 34 are arranged such that the follower part 34 is not in direct contact with the cam profile when the lever 30 is in the closed position. This latter embodiment has the advantage of limiting the friction of the follower part 34 on the cam profile and thus of limiting the wear of these two parts and the quantity of energy necessary for the rotation of the wheel 40.

[0064] To ensure the compactness of the device 1 according to the invention, the lateral size of each jaw is advantageously equivalent to 110% to 120% of that of a compressed flexible tube when said jaws are in the closed position. In the case where said jaw is intended to receive more than one flexible tube, the lateral size of said jaw is adapted accordingly.

[0065] Preferably, the flexible tubes used in the context of the present invention have a thickness of between 0.5 mm and 10 mm.

[0066] Preferably, the clamps 20 of a set of clamps 20 will be arranged parallel to each other. More particularly, the longitudinal axes of each lever 30 of a set of clamps 20 will be parallel to each other.

[0067] Even more preferably, the clamps 20 of the same set of clamps 20 are arranged side by side, separated from each other by a space necessary to avoid friction between said clamps 20 when their jaws are closed.

[0068] Preferably, all of the second jaws 22 of the same set of clamps 20 are arranged on the surface of the same plane. In order to facilitate the positioning of the flexible tubes between the jaws of two contiguous clamps, it is advantageous to alternately arrange the second jaws 22 of the same set of clamps 20, on two lines spaced at least the width of a flexible tube. Thus, said first and second jaws 21, 22 of two adjacent clamps 20 are arranged in a staggered pattern. In the embodiment where the offset of the jaw 21 is achieved via a blade 32 extending from the arm 31, the fact that said first and second jaws 21, 22 of two adjacent clamps 20 are arranged in a staggered pattern means that the two blades 32 of two adjacent clamps are arranged alternately on two lines spaced at least the width of a flexible tube.

[0069] Preferably, the wheels 40 of the wheel assembly 40 have an identical size and shape. Said wheels 40 are associated in rotation around the same axis A. By "solidarily" is meant the fact that all the wheels 40 of the same wheel assembly 40 rotate simultaneously around the axis a.

[0070] According to an even more preferred embodiment, the wheels 40 of the same set of wheels 40 are arranged so that the portion of the cam profile comprising the ascending, plateau and descending zones (corresponding to the opening phase of the jaws) is offset from one wheel to the other. Even more preferably, said portion occupies a different angular position for each wheel 40 of the set of wheels 40. It is thus understood that an angular position of the set of wheels 40 corresponds to the opening of a single jaw of the set of clamps 20.

[0071] Preferably, the set of wheels 40 is associated with a motor 60 capable of driving said set of wheels 40 in rotation around the axis A. Even more preferably, a motor with determination of the angular position 60 and most preferably a servomotor. The constant knowledge of the angular position of the motor 60 makes it possible to automatically deduce the position of the set of wheels 40 and consequently the position of each of the jaws of the set of clamps 20.

[0072] The association between the motor 60 and the wheel assembly 40 can be achieved by any means known to those skilled in the art. Among these, mention may in particular be made of gears, belts and direct coupling on the shaft of the wheel assembly 40.

[0073] The device 1 according to the invention advantageously comprises a chassis 70 on which the motor 60, the set of wheels 40 and the set of clamps 20 will be fixed.

[0074] Preferably, said chassis forms an enclosure of substantially parallelepiped shape.

[0075] Preferably, the motor 60 and the set of wheels 40 are arranged inside the chassis 70.

[0076] Even more preferably, the set of clamps 20 is partially arranged inside the chassis 70.

[0077] Most preferably, said chassis 70 comprises an opening 71 allowing access to the jaws 21, 22 of each of the clamps of the set of clamps 20. Advantageously, said jaws 21, 22 are arranged outside the chassis 70.

[0078] This latter embodiment allows the user to easily access the jaws to place the flexible tubes there without having to open the chassis 70.

[0079] The chassis 70 may preferably comprise an external surface 72 on either side of the opening 71 allowing access to the jaws 21, 22. This surface may advantageously receive guides 73 intended to block each of the flexible tubes on either side of its respective jaw.

[0080] The present invention also relates to a cell culture automaton characterized in that it comprises at least one device 1 according to the invention.

[0081] In addition to the device 1 according to the invention, the cell culture automaton according to the invention may comprise an electronic system controlling in particular the rotation of the motor 60 according to the invention. Furthermore, said electronic system may also control the operation of the pump when the latter is integrated into the cell culture automaton according to the invention.

[0082] A mode of use is described below. The user places in a temperature and gas regulated oven, a number n of culture dishes, a device 1 according to the invention under the control of an electronic system, a bottle containing culture medium, a container intended to collect the used media or cells.

[0083] A first tube is arranged between the culture medium bottle and the inlet of a disposable intermediate piece. The outlets of the intermediate piece are connected to N flexible tubes each connected at their other end to each of the N culture boxes.

[0084] Each flexible tube is arranged between the guides 73 and then passed between the jaws of a clamp 20. To do this, the user acts on the tab 33 to move the lever 30 and open the jaw. Alternatively, the user can program the sequential opening of the jaws via the electronic system. Once the flexible tube is in place, the jaw returns to the closed position under the effect of the elastic means. The flexible tube is thus closed and the passage of the culture medium blocked.

[0085] The same operation is carried out to connect the culture boxes to the enclosure intended to collect the waste.

[0086] The electronic system comprises software that manages the rotation of the wheel assembly 40 around its rotation axis A. Each time it is necessary to add or remove medium from inside a culture dish, the electronic system will actuate the motor 60 so that the latter drives the wheel assembly 40 in rotation. The rotation is carried out until the wheel 40, triggering the lever 30 of the clamp 20 associated with the flexible tube, is positioned so that the portion of the cam profile triggering the opening of the clamp 20 interacts with the follower part 34. Once the follower part is opposite the tray area, the rotation of the wheel assembly 40 is stopped for the period necessary for the addition of culture medium. The passage of the culture medium is triggered by the start of a pump by the electronic system electronics.

[0087] The feeding of the different culture boxes is thus carried out successively by rotation of the set of wheels 40. A complete rotation of this set of wheels 40 allows the successive opening of all the clamps 20 and therefore the feeding of all the culture boxes. The same applies to the extraction of the used medium.

Claims

Claims

1. Device (1) for controlling the passage of a liquid in a series of flexible tubes, comprising: - a set of clamps (20), each being capable of clamping a flexible tube, and comprising a first jaw (21) arranged on a lever (30) and a second jaw (22) forming a jaw with said first jaw (21), each lever (30) being pivotally mounted between an open position in which said clamp (20) is open and a closed position in which said clamp (20) is closed, and - a set of wheels (40) mounted integrally in rotation about the same axis A, each of said wheels (40) forming a cam and being associated with one of said levers (30), the rotation of each wheel (40) allows the passage of said lever (30) from its closed position to its open position and vice versa characterized in that it comprises a first stop (51) limiting the movement of said lever in the open position and a second stop (52) limiting the movement in closed position.

2. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to the preceding claim, characterized in that each lever (30) is forced into the closed position via an elastic means.

3. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to the preceding claim, characterized in that said elastic means is a spring.

4. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that said second jaw (22) is fixed

5. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that said first and second jaws (21, 22) of two contiguous clamps (20) are arranged in a staggered manner.

6. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of claims 1 to 5, characterized in that each second stop (52) is arranged so that said first and second jaws (21, 22) are not in contact in the closed position.

7. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that said first (21) and second jaws (22) are separated by a space equivalent to the thickness of a flexible tube when it is crushed.

8. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of claims 1 to 7, characterized in that each second stop (52) is arranged such that each lever (30) is not in contact with the cam path when said lever (30) is in the closed position.

9. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that each lever (30) comprises an arm (31) which extends perpendicular to the axis of rotation A.

10. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that each lever (30) is pivotally mounted around an axis B parallel to the axis of rotation A.

11. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that each lever (30) comprises an arm (31) from which extends perpendicularly an L-shaped blade (tongue) (33).

12. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that each lever (30) comprises at its distal end a tab (33) intended to allow the opening of the clamp by a user.

13. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that each wheel (40) comprises two parallel flat faces delimiting a cam profile comprising a neutral zone, an ascending zone, a plateau zone and a descending zone.

14. Device (1) for controlling the passage of a liquid in a series of flexible tubes according to one of the preceding claims, characterized in that said set of wheels (40) is associated with a motor with determination of the angular position (60) capable of driving said set of wheels (40) in rotation around the axis A.

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