Filtering device for suspended mesoscopic cellular objects

Abstract

The invention relates to a filtration device (1) for suspended mesoscopic cellular objects (MSCOs) comprising a body (2) having at least one first port (5) and a second port (6), and a filter membrane (3) extending along a plane P within the body (2), thus defining two compartments and capable of preventing the passage of MSCOs from one compartment to the other. The first port (5) is intended for the injection and withdrawal of MSCOs immersed in a soaking solution and / or a rinsing solution, and the second port (6) is intended for the withdrawal and injection of the soaking solution and / or the rinsing solution. According to the invention, injection into the first port (5) is carried out along a direction D not substantially parallel to the plane P of the filter membrane (3). Figure for the abstract: Fig. 1

Description

Title of the invention: Device for filtering mesoscopic cellular objects in suspension

[0001] The present invention relates to the technical field of devices for filtering mesoscopic cellular objects in suspension.

[0002] Mesoscopic cellular objects in suspension, hereinafter referred to as OCMS, comprise a set of objects whose dimensions vary between 5 pm and 5 mm, which are in suspension and which comprise at least one eukaryotic cell, encapsulated or not. For example, OCSM comprise human cells in suspension, microtissues, spheroids, cell aggregates, “embryoid bodies”, encapsulated or not.

[0003] In the above field, OCMS filtration modules are known comprising a body comprising a filter membrane intended to retain the OCMS but for example capable of allowing the solution in which the OCMS are bathed to pass through.

[0004] Several filtration techniques exist, in particular frontal filtration, where the OCMS are injected normally at the plane of the filter membrane. This technique can cause the accumulation and compaction of the OCMS on the membrane which can prevent the good filtration of the OCMS, damage them and clog the filter.

[0005] Another filtration technique is tangential filtration, where the OCMS in solution are injected through a tube porous to the solution but not to the OCMS, in which the solution is drawn through the wall of the tube, while the OCMS pass through the tube. This tangential filtration technique makes it possible to minimize the risk of accumulation and compaction. The disadvantage of this technique is that it relies on filters that are complex to manufacture and that it hardly allows a complete rinsing of the OCMS. Indeed, there always remains a residual proportion of soaking solution after rinsing due to the fact that the OCMS are not confined in a compartment and multiple rinsing steps are required to achieve the desired rinsing solution concentration.

[0006] Another filtration technique is notably centrifugal filtration which does not use a membrane but the difference in density of the OCMS compared to the soaking / rinsing solution to separate OCMS and solution. This technique requires heavy equipment (centrifuge), expensive and complex to set up.

[0007] The present invention aims to overcome these drawbacks.

[0008] The invention therefore relates to a device for filtering suspended mesoscopic cellular objects (OCMS) comprising a body comprising at least a first port and a second port and a filter membrane extending along a plane P in the body, thus defining two compartments and capable of preventing the passage of OCMS from one compartment to another, the first port being intended for the injection and withdrawal of the OCMS bathed in a soaking solution and / or a rinsing solution and the second port being intended for the withdrawal and injection of the soaking solution and / or the rinsing solution.

[0009] According to the invention, the injection into the first port is carried out in a direction D not substantially orthogonal to the plane P of the filter membrane.

[0010] Advantageously, the injection into the first is a confined tangential of the OCMS and thus makes it possible not to create an accumulation or damage to the OCMS.

[0011] A tangential flow or vortex in the filtration device reduces damage to the OCMS by keeping the OCMS moving.

[0012] According to a characteristic of the invention, the direction D is parallel to the plane P.

[0013] According to one embodiment of the invention, the body comprises an inner casing with edges having a rounded shape and the useful section of the filter membrane has the shape of a circle or an ellipse. The implementation of rounded edges makes it possible to reduce the recesses inside the body of the device, thus limiting the accumulation of OCMS in the recesses and making it possible to avoid the loss of OCMS trapped in the device during the OCMS recovery phases. This also makes it possible to limit the dead volume and therefore the volume necessary to rinse the OCMS.

[0014] According to another embodiment of the invention, the body has a toric shape and the useful section of the filter membrane has the shape of a circle or an ellipse. Advantageously, the toric shape makes it possible to increase the swirl effect for the OCMS while greatly reducing the dead volume inside the body, thus limiting the volume necessary to rinse the OCMS.

[0015] According to a characteristic of the invention, any section perpendicular to the plane P of the compartment intended to receive the OCMS has a surface area strictly less than the useful surface area of ​​the filter membrane.

[0016] According to another characteristic of the invention, any section perpendicular to the plane P of the compartment intended to receive the OCMS has a surface area twice, five times and preferably ten times smaller than the useful surface area of ​​the filter membrane.

[0017] The tangential injection associated with such a difference in dimension makes it possible to inject the OCMS with a certain flow rate, promoting the swirl, while allowing the concentration of a certain number of OCMS inside the body of the filtration device and minimizing the dead volume.

[0018] According to a characteristic of the invention, the injection into the second port is also carried out in a non-orthogonal direction, and preferably parallel, to the plane P of the filter membrane. The second tangential injection makes it possible to promote swirling flows inside the body of the filtration device.

[0019] According to yet another characteristic of the invention, the soaking solution is a solution allowing cell culture. The OCSM are thus maintained in a favorable environment and are not degraded.

[0020] According to a characteristic of the invention, the soaking solution contains ions from the following list: calcium, iron, barium, strontium, copper, lead, aluminum, magnesium, preferably calcium, iron, barium. The addition of such ions is of great interest for the good performance of the OCMS.

[0021] According to another characteristic of the invention, the filter membrane comprises a microfabric covered with a fiber membrane. The implementation of such a filter membrane makes it possible to maintain control over the type of OCMS to be filtered.

[0022] According to one embodiment of the invention, the filter membrane is capable of allowing other OCMS of a different type contained in the soaking solution to pass through. This characteristic in particular allows filtering of a certain type of OCMS.

[0023] The invention also relates to a method for accumulating OCMS comprising the following steps: - Injection of OCMS bathed in the soaking solution into a first port of an OCMS filtration device according to the invention, filtration of the OCMS by the filter membrane and recovery of a portion of the soaking solution by the second port, - Injection of the soaking solution into the second port and recovery of the accumulated OCMS through the first port.

[0024] According to one embodiment of the invention, the OCMS accumulation process is carried out continuously. The OCMS can thus be collected as quickly as they are produced, thus minimizing the residence time in the soaking solution.

[0025] The invention also relates to a method for cleaning OCMS comprising the following steps: - Injection of OCMS bathed in a soaking solution into a first port of an OCMS filtration device according to the invention, filtration of the OCMS by the filter membrane and recovery of the soaking solution and the rinsing solution by the second port, - Injection of a rinsing solution into the first given port intended to replace the soaking solution and recovery of the two solutions and rinsing by the second port.

[0026] Advantageously, the tangential flow reduces damage to the OCMS by minimizing contact with the filter membrane during the cleaning process.

[0027] According to a characteristic of the invention, the cleaning method further comprises a step of injecting the rinsing solution into the second port and recovering the cleaned OCMS via the first port.

[0028] The invention also relates to a cell encapsulation system, the system comprising at least two containers, one of the containers comprising a cell solution and the other of the containers comprising a solution capable of gelling, an encapsulation device comprising a fluidic chip comprising main and secondary inlets each being connected to one of the containers via a distributor and capable of forming a concentric jet from the solutions supplied by the distributors, the encapsulation device being arranged so that the jet splits, at the outlet of the encapsulation device, into drops of which the outer layer is the solution capable of gelling and the core the cell solution, a gelling bath arranged downstream of the encapsulation device to collect the drops formed by the encapsulation device and arranged to cause gelling of the outer layer of each drop during its immersion in the bath,and a filtration device according to the invention.

[0029] Of course, the various features, variants and embodiments of the invention may be combined with each other in various combinations to the extent that they are not incompatible or mutually exclusive.

[0030] Furthermore, various other characteristics of the invention emerge from the appended description given with reference to the drawings which illustrate non-limiting embodiments of the invention and where:

[0031] [Fig-1] is a perspective view of an exemplary embodiment of a device for filtration according to the invention,

[0032] [Fig.2] is a schematic view of an exemplary embodiment according to the invention,

[0033] [Fig.3] is a perspective view of an exemplary embodiment of a filtration device according to the invention,

[0034] [Fig.4] is a half-sectional view of [Fig.3] along the section plane AA,

[0035] [Fig.5] is a half-sectional view of another exemplary embodiment of the invention, And

[0036] [Fig.6] is a schematic view of an exemplary embodiment with two filtration devices according to the invention.

[0037] It should be noted that in these figures the structural and / or functional elements common to the different variants may have the same references.

[0038] The invention aims to filter suspended mesoscopic cellular objects, known as OCMS.

[0039] For these purposes, the invention designated by the reference 1 as a whole and illustrated in [Fig.l] relates to a filtration device comprising a body 2 comprising a filter membrane 3 which extends in the body 2 along a plane P.

[0040] According to the invention, the body 2 of the filtration device 1 comprises an inner casing 4 having rounded edges.

[0041] According to one embodiment of the invention illustrated in particular in Figures 2 and 5, the inner casing 4 has the shape of a burger, that is to say a cylindrical shape with a rounded upper edge surface and a rounded lower edge surface.

[0042] According to the embodiment illustrated in Figures 3 and 4, the inner envelope 3 of the body 2 has the shape of a donut, that is to say a toric shape.

[0043] According to the embodiment illustrated in [Fig. 2], the filtration device 1 is symmetrical with respect to the filter membrane 3. Other non-symmetrical embodiments illustrated in Figures 3, 4 and 5 are also compatible with the invention. For example, the part intended to receive OCMS may comprise larger volume dimensions than the part on the other side of the filter membrane 3 or vice versa.

[0044] The filter membrane 3 extends along a plane P in the body 2 of the filtration device 1. The filter membrane 3 extends to the edges of the inner casing 4 of the body 2 of the filtration device 1. According to the illustrated embodiments, the useful section of the filter membrane 3 has the shape of a circle. According to other embodiments not illustrated, the useful section of the filter membrane 3 has the shape of an ellipse.

[0045] In order to retain the OCMS, the filter membrane 3 comprises a fiber membrane capable of preventing the passage of the OCMS.

[0046] The filter membrane 3 is therefore adapted according to the type of OCMS implemented.

[0047] Mesoscopic cellular objects in suspension (MCSOs) refer to single cells or microtissues, but also to cells encapsulated in alginate microcompartments.

[0048] The OCMS are immersed in a soaking solution. Advantageously, the soaking solution allows cell culture. For example, the OCMS are cells encapsulated in alginate microcompartments and the soaking solution contains Calcium Chloride CaCl2.

[0049] According to one embodiment of the invention, the filter membrane 3 comprises a microfabric covered with a fiber membrane. The fiber membrane contains a material of the wide mesh textile type (mesh), nylon, or PET. Other filter materials, OCMS, are used in the context of the invention.

[0050] According to one embodiment of the invention, the filter membrane 3 is capable of filtering at least one type of OCSM and is also capable of allowing another type of OCMS to pass through. The mesh can thus be adapted according to the dimensions of the OCMS to be filtered.

[0051] The body 2 of the filtration device 1 comprises a first port 5 intended for tangential injection of the OCSM bathed in a soaking solution.

[0052] The first port 5 comprises for its purposes a cylindrical shape which extends substantially in a direction D belonging to a plane P' parallel to the plane P of the filter membrane 3 as shown in [Fig.l].

[0053] The second port 6 is located in the second part of the body 2, on the other side of the filter membrane 3. According to the embodiment illustrated in Figures 1 and 2, the second port 6 does not allow tangential injection into the body 2 of the filtration device 1. According to the embodiment illustrated in particular in Figures 3, 4 and 5, the second port 6 is also suitable for tangential injection into the body 2 of the filtration device 1.

[0054] According to the embodiment illustrated in [Fig.l], the first port 5 comprises an inlet 7 and an injection channel 8.

[0055] The injection channel 8 allows tangential injection into the inner casing 4 of the body 2 of the filtration device 1.

[0056] According to other embodiments not illustrated, the body 2 of the filtration device 1 comprises more than two inlet and outlet ports thus allowing the injection of different types of solution or OCMS on each side of the filter membrane 3.

[0057] The filtration device 1 according to the invention is used for different functions.

[0058] The filtration device 1 is notably used for various processes such as OCSM accumulation and OCMS cleanup.

[0059] A first use of the filtration device 1 relates to a method of accumulating OCMS.

[0060] A first step of the OCMS accumulation method consists of injecting OCMS bathed in the soaking solution into the first port 5 of the OCMS filtration device 1 according to the invention. The injection is carried out according to a flow rate Q1 for a time TL. For example, the flow rate Q1 is 130 mL / min and the time T1 is 45 s. The OCMS are stopped by the filter membrane 3. A portion of the soaking solution is discharged through the second port 6 of the filtration device 1.

[0061] A second step in the OCMS accumulation process consists of a flow reversal. To do this, the previously evacuated soaking solution is injected into the second port 6.

[0062] The OCMS accumulated in the filtration device 1 during the first step of the OCMS accumulation process are then recovered via the first port 5.

[0063] Such a method of accumulating OCMS using the filtration device 1 according to the invention can advantageously be continuous by repeating the two steps described.

[0064] A second use of the filtration device 1 according to the invention relates to a method for cleaning OCMS.

[0065] A first step of the OCMS cleaning method consists of injecting OCMS bathed in a soaking solution into the first port 5 of the filtration device 1 at a flow rate Q2, and filtering the OCMS by the filter membrane 3 and recovering a portion of the soaking solution through the second port 6 during a time T2. For example, the flow rate Q2 is 260 mL / min and the time T2 is 15 s.

[0066] A second step of the OCMS cleaning method consists of injecting a rinsing solution into the first port 5 at a flow rate Q3 for a given time T3, the rinsing solution then replacing the soaking solution and recovering the soaking solution mixed with the rinsing solution and then the rinsing solution alone via the second port 6. For example, the flow rate Q3 is 250 ml / min and the time T3 is 20 s.

[0067] A third step consists of reversing the flow with the injection into the second port 6 of the rinsing solution at a flow rate Q4 and the recovery of the cleaned OCMS by the first port 5 for a time T4. For example, the flow rate Q4 is 250 mL / min and the time T4 is 10 s.

[0068] Such an OCMS cleaning method using the filtration device 1 according to the invention can advantageously be continuous by repeating the three steps described.

[0069] [Fig. 6] illustrates the implementation of the two methods described above in series. A first filtration device 1 is used for an OCMS accumulation process and a second filtration device is used for an OCMS cleaning process.

[0070] Two filtration devices 1 are used to accumulate the produced OCMS and transfer them, and to wash the OCMS with a rinsing solution and to collect the OCMS in a closed single-use system. To do this, a flow valve 10 is interposed between the two respective first ports of the two filtration devices 1. And the steps described above can take place continuously.

[0071] The first port 5 of a first filtration device 1 is connected to the first port 5 of a second filtration device 1 in order to be able to directly inject OCSM accumulated by the first filtration device 1 and to clean them using the second filtration device 1.

[0072] Other forms of architecture may be formed according to the use of the invention.

[0073] Of course, various other modifications may be made to the invention within the scope of the appended claims.

Claims

Claims

1. Filtration device (1) for suspended mesoscopic cellular objects (OCMS) comprising a body (2) comprising at least a first port (5) and a second port (6) and a filter membrane (3) extending along a plane P in the body (2) thus defining two compartments and capable of preventing the passage of the OCMS from one compartment to the other, the first port (5) being intended for the injection and withdrawal of the OCMS bathed in a soaking solution and / or a rinsing solution and the second port (6) being intended for the withdrawal and injection of the soaking solution and / or the rinsing solution, characterized in that the injection into the first port (5) is carried out along a direction D not substantially orthogonal to the plane P of the filter membrane (3).

2. Filtration device (1) according to the preceding claim in which the direction D is parallel to the plane P.

3. Filtration device (1) according to one of the preceding claims in which the body (2) comprises an inner casing with edges having a rounded shape and the useful section of the filter membrane (3) has the shape of a circle or an ellipse.

4. Filtration device (1) according to one of the preceding claims in which any section perpendicular to the plane P of the compartment intended to receive the OCMS has a surface area strictly less than the useful surface area of ​​the filter membrane (3).

5. Filtration device (1) according to the preceding claim in which any section perpendicular to the plane P of the compartment intended to receive the OCMS has a surface area twice, five times and preferably ten times smaller than the useful surface area of ​​the filter membrane.

6. Filtration device (1) according to one of the preceding claims in which the body (2) has a toric shape and the useful section of the filter membrane (3) has the shape of a circle or an ellipse.

7. Filtration device (1) according to one of the preceding claims in which the injection into the second port (6) is also carried out in a direction which is non-orthogonal, and preferably parallel, to the plane P of the filter membrane (3).

8. Filtration device (1) according to one of the preceding claims in which the soaking solution is a solution allowing cell culture.

9. Filtration device (1) according to the preceding claim in which the soaking solution contains ions from the following list: calcium, iron, barium, strontium, copper, lead, aluminum, magnesium, preferably calcium, iron, barium.

10. Filtration device (1) according to one of the preceding claims in which the filter membrane (3) comprises a microfabric covered with a fibrous membrane.

11. Filtration device (1) according to one of the preceding claims in which the filter membrane (3) is capable of allowing other OCMS of a different type contained in the soaking solution to pass through.

12. Method for accumulating OCMS comprising the following steps: - Injecting OCMS bathed in the soaking solution into a first port (5) of a filtration device (1) according to one of claims 1 to 11, filtering the OCMS by the filter membrane (3) and recovering a portion of the soaking solution through the second port (6), - Injecting the soaking solution into the second port (6) and recovering the accumulated OCMS through the first port (5).

13. Method for cleaning OCMS comprising the following steps: - Injecting OCMS bathed in a soaking solution into a first port (5) of a filtration device (1) according to one of claims 1 to 11, filtering the OCMS by the filter membrane (3) and recovering the soaking solution and the rinsing solution through the second port (6), - Injecting a rinsing solution into the first given port (5) intended to replace the soaking solution and recovering both solutions and rinsing through the second port (6).

14. A method of cleaning OCMS according to the preceding claim further comprising a step of injecting the rinsing solution into the second port (6) and recovering the cleaned OCMS through the first port (5).

15. Cell encapsulation system, the system comprising at least: - two containers, one of the containers comprising a cell solution and the other of the containers comprising a solution capable of gelling, - an encapsulation device comprising a fluidic chip comprising main and secondary inlets each being connected to one of the containers via a distributor and capable of forming a concentric jet from the solutions supplied by the distributors, the encapsulation device being arranged so that the jet splits, at the outlet of the encapsulation device, into drops of which the outer layer is the solution capable of gelling and the core the cell solution, - a gelling bath arranged downstream of the encapsulation device to collect the drops formed by the encapsulation device and arranged to cause gelling of the outer layer of each drop when it is immersed in the bath, and - a filtration device (1) according to one of claims 1 to 11.

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