Bioprocessing chamber having a frustoconical cross-section

Abstract

The invention relates to a process chamber (100) which is shaped concentrically around a central axis (101). The process chamber (100) comprises a first body section (1) which is frustoconical and has a first base side (11) and a second base side (12). The second base side (12) opens into a second body section (2). The second body section extends along the central axis (101), a first end of the second body section (2) pointing away from the first body section (1). The first end of the second body section (2) opens into a third body section (3). The third body section extends along the central axis (101), a first end of the third body section (3) pointing in a direction away from the second body section (2), wherein the first end of the third body section (3) tapers into a second end (30) of the process chamber (100).

Description

Technical Field

[0001] The invention relates in a first aspect to a processing chamber having a truncated first body section, in a second aspect to a kit including a processing chamber, in a third aspect to an apparatus suitable for receiving a processing chamber, in a fourth aspect to a method for culturing and processing cells in a processing chamber, and in a fifth aspect to the use of a processing chamber. Background Technology

[0002] Processing chambers are well-known in the field of cell culture and processing. In particular, document WO2019 / 234033 describes an apparatus and method for the automated production of genetically engineered cells from biological fluids.

[0003] This document describes a hollow processing chamber consisting of a hemispherical top portion, with the circular portion of the hemisphere facing upwards and having an opening at the very top. The edges of the hemisphere form a vertical angle with the hemisphere itself.

[0004] In any case, the spherical shape of the processing chamber cannot provide a large incubator volume for cell processing compared to the overall volume of the chamber. Furthermore, due to its spherical surface, the inner surface of the hemisphere is not very advantageous for incubating adherent cells. Summary of the Invention

[0005] Therefore, the problem to be solved by the present invention is to provide a processing chamber that provides a better total volume to incubator volume ratio and is more suitable for centrifugation and cell incubation, especially for incubating adherent cells.

[0006] The “processing chamber” is advantageously a centrifugal separation chamber.

[0007] The “processing room” is advantageously a bioreactor.

[0008] The term "chamber" advantageously refers to a hollow container or tank that can be filled with liquid.

[0009] The orientation of "vertical" is advantageously defined by the direction of local gravity.

[0010] The orientation "horizontal" advantageously refers to the direction perpendicular to the vertical direction.

[0011] This problem is solved by the processing chamber according to the first aspect of the invention.

[0012] The processing chamber is concentrically formed around a central axis and includes a first main section, a second main section, and a third main section, wherein, along the central axis, the first main section is followed by the second main section, and the second main section is followed by the third main section. Specifically, it should be clarified that the central axis is the central axis of the processing chamber itself.

[0013] The first main body section has a truncated conical shape with a first base side and a second base side. Advantageously, according to the definition of a truncated conical shape, the first base side and the second base side of the first main body section can be arranged parallel to each other, especially substantially parallel. This should also include the first base side and the second base side of the first main body section that are slightly misaligned and not perfectly parallel due to manufacturing problems or similar issues.

[0014] The diameter of the first base side of the first main body section is smaller than the diameter of the second base side of the first main body section, and the first base side forms the first end of the processing chamber in the axial direction of the central axis.

[0015] The second base of the first main body section extends into the second main body section. The second main body section extends along a central axis, wherein the first end of the second main body section points away from the first main body section, and the second end of the second main body section is adjacent to the first main body section. Advantageously, the diameter of the second main body section at the first end is smaller than the diameter of the second main body section at the second end.

[0016] Advantageously, an angle or edge is formed between the first body section and the second body section. This angle or edge is formed between the lateral inner surface of the first body section (the inner surface between the first base side and the second base side of the first body section, particularly the inner shell surface between the first base side and the second base side of the first body section) and the lateral inner surface of the second body section (the surface between the first end and the second end of the second body section).

[0017] The first end of the second main section leads into the conical third main section.

[0018] The third main section extends along the central axis in a direction pointing away from the second main section, wherein the first end of the third main section points away from the second main section. The first end of the third main section tapers to the second end of the processing chamber. Advantageously, the diameter of the second end of the processing chamber is smaller than the diameter at the first end of the second main section.

[0019] Advantageously, tapering into the second end of the processing chamber means that the third body section tapers until it ends at the tip or until an opening is left at the second end of the processing chamber.

[0020] Advantageously, the third main body section has a conical shape with an opening at the tip, and in particular, it has a truncated or frustum-like shape.

[0021] Advantageously, the processing chamber, comprising a first main body segment, a second main body segment, and a third main body segment, has a favorable shape for incubating cells. In particular, since the volume of the substantially truncated first main body segment is large compared to the total volume of the processing chamber, a larger volume of liquid can be centrifuged and incubated compared to processing chambers described in the prior art.

[0022] In another advantageous embodiment of the invention, the slope of the second body section about the central axis is greater than the slope of the third body section about the central axis.

[0023] Advantageously, the term "tilt height" here refers to the height of an object (e.g., a truncated body), which is the distance measured along the lateral plane from the first base to the vertex or the second base and along the center of the lateral plane.

[0024] In another advantageous embodiment of the invention, the second base side of the first body section extends into the truncated second body section.

[0025] Advantageously, the diameter of the first base side of the second main body section closer to the first main body section is greater than the diameter of the second base side of the second main body section away from the first main body section.

[0026] In another advantageous embodiment of the invention, the height h of the truncated second body segment f for h f ≤10mm, specifically h f ≤5mm.

[0027] Further advantageously, the second main body section is designed such that its surface line follows an elliptical, substantially elliptical, or circular contour into the surface line of the third main body section. Specifically, the surface line of the second main body section refers to the shortest line traveling along an inclined height between the second end of the second main body section and the first end of the second main body section. More specifically, the surface line of the third main body section refers to the shortest line between the second end of the third main body section and the first end of the third main body section in the direction of the central axis.

[0028] In another advantageous embodiment of the invention, the minimum angle α between the tilt height of the second main body section and the second base side is α≤20°, particularly α≤10°, particularly α≤5°.

[0029] In another advantageous embodiment of the invention, the processing chamber includes a first opening for flow exchange at a first end and / or a second opening for flow exchange at a second end.

[0030] In another advantageous embodiment of the invention, at least the inner shell surface or lateral inner shell surface of the first body section is coated with a bioactive coating. This means that at least the inner wall of the first body section is coated, particularly to better facilitate cell adhesion.

[0031] A second aspect of the invention relates to a kit comprising a processing chamber according to a first aspect of the invention. The processing chamber in the kit is connected to a first flow stator and / or a second flow stator. Furthermore, the kit includes a tubing assembly connectable to the first flow stator and / or the second flow stator, and adapted, for example, in a device, in the intended use of the kit, to be connected to at least one peristaltic pump head and at least one pinch valve. Additionally, the kit includes at least one reagent or product bag or bottle adapted to be connected to the tubing assembly.

[0032] A third aspect of the invention relates to an apparatus adapted to receive or include a processing chamber or a kit having a processing chamber. The apparatus is adapted to prepare gene-transformed cells from a biological fluid. The apparatus may include an operating unit adapted to receive the processing chamber according to the first aspect. Further advantageously, the apparatus may be adapted to orient the processing chamber about a rotational axis of the operating unit, particularly to orient the central axis of the processing chamber in a horizontal or vertical direction relative to the ground.

[0033] A fourth aspect of the present invention relates to a method for culturing and processing cells in a processing chamber according to a first aspect of the present invention. The method includes the following steps: Bring the processing chamber in so that the central axis is vertically aligned to allow the cells to be centrifuged, and / or Bring the processing chamber in so that the central axis is horizontally aligned with the position for incubating cells (especially suspended cells), and / or The operating unit is brought into position such that the surface line of the first main body segment is horizontally aligned for incubating cells (particularly adherent cells). Specifically, the surface line of the first main body segment refers to the shortest line traveling along the slope between the first and second basal sides of the first main body segment. More specifically, horizontal alignment of the surface line of the first main body segment means aligning the surface line along a horizontal plane.

[0034] Advantageously, the method is used to prepare gene-transformed cells from biological fluids using a processing chamber according to a first aspect of the invention and an apparatus according to a third aspect of the invention.

[0035] Further advantageously, the method therefore includes the step of rotating the operating unit of the device to a position where the central axis of the processing chamber is in a vertical position. Additionally, the method includes one of the following steps: rotating the operating unit to a position where the central axis of the processing chamber is in a horizontal position, or rotating the operating unit to a position such that the surface line of the first body section is horizontally aligned for incubating cells.

[0036] Bringing the processing chamber to a horizontal position means arranging the central axis horizontally, which can effectively incubate suspended cells.

[0037] Positioning the central axis of the processing chamber at a slight inclination relative to the horizontal plane means that the surface line of the first main section is horizontally aligned, which allows for effective incubation of adherent cells. In this position, cells can effectively adhere to the lateral inner surface of the first main section, particularly to the lateral inner surface of the first main section which is substantially horizontally aligned in this position of the processing chamber.

[0038] Advantageously, particularly regarding the handling of adherent cells, this method allows switching between three positions: a vertical central axis, a horizontal central axis, and a tilted central axis that aligns the surface line of the first main body segment horizontally. This is one of the advantages of the method, which allows for efficient centrifugation and incubation of biological cells while, for example, exchanging fluids.

[0039] Advantageously, the step of bringing the processing chamber to a vertical position with the central axis vertically aligned is a method step suitable for subjecting the liquid (cell suspension) in the processing chamber to centrifugation. Advantageously, the cell suspension can be centrifuged such that cell clusters form on the lateral inner surface near the first main section, and on the lateral inner surface between the first and second main sections, particularly at or near the edges, particularly formed between the surfaces and the angle formed between the first and second main sections. Further advantageously, when centrifugation is slowed or stopped, the supernatant or any other fluid can be collected at the bottom of the processing chamber. The supernatant or any other fluid can be removed from the processing chamber through a second opening in the processing chamber.

[0040] Further advantageously, after the centrifugation step, the processing chamber is brought to a position where the central axis is horizontally aligned. A liquid or solution can then be added to the processing chamber. The cell clumps can now be suspended in the solution by stirring. The suspended cells can then be further incubated to promote cell division. The advantage of the processing chamber now lies in the increased volume and increased lateral internal surface area resulting from its innovative shape.

[0041] In another advantageous embodiment of the invention, the method further includes the step of exchanging the liquid or solution in the processing chamber via a first opening and / or a second opening. Advantageously, the liquid or solution in the processing chamber is exchangeable only when the processing chamber is in a static position.

[0042] In another advantageous embodiment of the method, particularly for the centrifugation and incubation of suspended cells, the method may further include the following steps: Especially when the processing chamber is in a position where the central axis is vertically aligned, add (cell) suspension to the processing chamber, and / or The (cell) suspension is centrifuged by rotating the processing chamber around its central axis, particularly wherein cell clusters are formed on the lateral inner surface near the first main body segment, between the first and second main body segments, at the edges between the first and second main body segments, and / or When centrifugation slows down or stops, collect the supernatant at the bottom of the processing chamber (in the third main section), and / or Advantageously, after the centrifugation step, the processing chamber is brought to a horizontal position (with the central axis in a horizontal position) to allow any liquid or solution to be added to the processing chamber, and / or In particular, cell clusters are suspended by stirring, and / or While the processing chamber is in a horizontal position, the suspended cells are further incubated to promote cell division.

[0043] In another advantageous embodiment of the method, particularly for the centrifugation and incubation of adherent cells, the method may further include the following steps: The processing chamber is brought into a position where the surface line of the first main section is horizontally aligned, and a suspension of adherent cells, particularly from the growth medium, is added to the processing chamber, and / or Especially during the rotation of the processing chamber, cells form an adherent layer on the lateral inner surface of the first main segment, and / or When the treatment chamber is positioned horizontally to the surface line of the first main section (the surface line of the first main section is horizontally aligned), add the growth medium to the treatment chamber, and / or When the processing chamber is in a vertical position (central axis in a vertical position), the growth medium is collected and removed, while the adherent cell layer remains on the lateral inner surface of the first main section, and / or Especially after cell growth, particularly when the treatment chamber is positioned at the level of the surface line of the first body segment, a trypsinizing agent is added to the treatment chamber, wherein the trypsinizing agent can release cells attached to the laterally inner surface of the first body segment, and / or The trypsin solution and trypsin reagent were removed separately from the processing chamber along with the suspended cells for further use.

[0044] Various embodiments of the method according to the fourth aspect are advantageously used for closed and automated cell processing. In particular, automated cell processing is provided if the method is operated on an apparatus according to the third aspect of the invention. The special shape of the processing chamber allows different method steps to be run in all steps using the same processing chamber in a sterile and closed environment. Therefore, the result is more efficient and safer cell culture and processing, as all necessary steps of centrifugation and growth can be performed in the same processing chamber.

[0045] The fifth aspect of the invention relates to the use of a processing chamber according to the first aspect of the invention for preparing gene-transformed cells from a biological fluid by a method according to the fourth aspect of the invention.

[0046] Other advantageous embodiments are set forth in the dependent claims and in the description below. Attached Figure Description

[0047] The invention will be better understood from the following detailed description, and objectives other than those set forth above will become apparent. This description is made with reference to the accompanying drawings, in which: Figure 1a to Figure 1c Three different views of the processing chamber according to an embodiment of the first aspect of the present invention are shown; Figures 1d to 1f Showing from Figure 1c Detail A; Figures 2a to 2c Three different views of a processing chamber according to another embodiment of the first aspect of the present invention are shown; Figures 2d to 2e Detail B of Figure 2 is shown; Figure 3 illustrates another embodiment of the processing chamber according to the first aspect of the present invention; Figure 4 A kit illustrating an embodiment of the second aspect of the present invention is shown; Figure 5 shows the device equipped with the kit, in which the processing chamber aligns its central axis along the vertical plane and the horizontal plane.

[0048] Figure 6The steps of an embodiment of the method according to the fourth aspect of the invention are shown using a first embodiment of the processing chamber; Figure 7 The steps of an embodiment of a method for processing suspended cells according to the fourth aspect of the invention are shown using a second embodiment of the processing chamber; and Figure 8 The steps of another embodiment of the method for treating adherent cells according to the fourth aspect of the present invention are shown using one embodiment of the processing chamber. Detailed Implementation

[0049] Figures 1a to 1c Three different views of a first embodiment of the processing chamber 100 are shown.

[0050] Figure 1a A three-dimensional view of a processing chamber 100 concentrically formed around a central axis 101 is shown. The processing chamber 100 includes a first main body section 1 that is substantially truncated (particularly truncated conical) and has a first base side 11 and a second base side 12. The first base side 11 and the second base side 12 of the first main body section 1 are advantageously parallel to each other or substantially parallel to each other.

[0051] The first base side 11 of the first main body section 1 has a smaller diameter than the second base side 12 of the first main body section 1, and forms the first end 10 of the processing chamber 100 in the axial direction of the central axis 101. The second base side 12 of the first main body section opens into the second main body section 2. The second main body section 2 extends along the central axis 101, wherein the first end of the second main body section 2 points away from the first main body section 1. The first end of the second main body section 2 opens into the third main body section 3. The third main body section 3 extends along the central axis 101, wherein the first end of the third main body section 3 points away from the second main body section 2. The first end of the third main body section 3 gradually tapers into the second end 30 of the processing chamber.

[0052] Advantageously, such as from Figure 1b As can be seen, the second main body segment 2 can be truncated, having a first base side and a second base side. The first base side of the second main body segment 2 is closer to the first main body segment 1 and is larger in diameter than the second base side of the second main body segment 2.

[0053] Advantageously, such as Figure 1d As shown, the height h of the truncated second main body section 2 f for h f ≤10mm, specifically h f ≤5mm.

[0054] Advantageously, the taper at the first end of the third main body section 3 may terminate at a tip or at an opening, such as Figures 1a to 1c As shown.

[0055] In a further advantageous embodiment, the slope "a" of the inclination height of the second main body section about the central axis 101 is greater than the slope "b" of the inclination height of the third main body section about the central axis 101, such as... Figure 1e As shown.

[0056] Further advantageously, such as Figure 1f As shown, the minimum angle α between the tilt height of the second main body section 2 and the second base side 12 of the first main body section 1 is α≤20°, specifically α≤10°, and specifically α≤5°.

[0057] Further advantageously, such as from Figures 1a to 1c As can be seen, the processing chamber 101 includes a first opening 13 for flow exchange at the first end 10 and / or a second opening 31 for flow exchange at the second end 30.

[0058] Figures 2a to 2c Three different views of a second embodiment of the processing chamber 100 are shown.

[0059] Figures 2a to 2c Another embodiment of the processing chamber 100 is shown. The second embodiment is similar to... Figures 1a to 1c The difference in the first embodiment shown is that the surface line of the second body section 2 follows a generally elliptical or circular outline in the surface line of the third body section 3.

[0060] In an advantageous second embodiment, the slope "a" of the second main body section's inclination height about the central axis 101 is greater than the slope "b" of the third main body section's inclination height about the central axis 101, such as... Figure 2d As shown.

[0061] Further advantageously, such as Figure 2e As shown, the minimum angle α between the tilt height of the second main body section 2 and the second base side 12 of the first main body section 1 is α≤20°, specifically α≤10°, and specifically α≤5°.

[0062] Further advantageously, such as from Figures 2a to 2c As can be seen, the processing chamber 101 includes a first opening 13 for flow exchange at a first end 10 and / or a second opening 31 for flow exchange at a second end 30.

[0063] In a further advantageous embodiment of the processing chamber, the processing chamber further includes a first flow stator at a first end 10 and / or a second flow stator at a second end 30.

[0064] Further advantageously, the treatment chamber may include a bioactive coating or any other coating on the lateral inner surface of the first body section 1.

[0065] like Figure 3a and Figure 3b As shown, in a further advantageous embodiment of the processing chamber, the processing chamber further includes a first flow stator 40 at a first end 10 and / or a second flow stator 50 at a second end 30. Figure 3a and Figure 3b In the middle, it is shown that according to Figures 1a to 1c An embodiment of the processing chamber 100 has a first flow stator 40 and / or a second flow stator 50. However, the first flow stator 40 and / or the second flow stator 50 may also be included according to... Figures 2a to 2c In the processing chamber 100 of the illustrated embodiment or any other embodiment, the processing chamber 100 is connected to a static element (the corresponding first flow stator 40 or second flow stator 50) by means of a bearing 42 as a rotating element, and the fluid contents are sealed by means of a rotary seal 41. When the stator is not present at the first opening 13 or the second opening 31, the corresponding first opening 13 and / or second opening 31 includes a filter 43 to allow sterile air to pass through the corresponding first opening 13 and / or second opening 31.

[0066] Further advantageously, the treatment chamber may include a bioactive coating or any other coating on the lateral inner surface of the first main body section 1.

[0067] Figure 4 An embodiment of a kit 200 including a processing chamber 100 is shown. The processing chamber 100 in kit 200 is connected to a first flow stator 40 and / or a second flow stator 50. Furthermore, kit 200 includes a tubing assembly 60 connectable to the first flow stator 40 and / or the second flow stator 50, and adapted, for example, to connect to at least one peristaltic pump head and at least one pinch valve in the intended use of the tubing assembly, in device 300 (as depicted in FIG. 5). Additionally, kit 200 includes at least one reactant or product bag 61 or bottle adapted to connect to the tubing assembly 60.

[0068] Figure 5a and Figure 5b An apparatus 300 according to an embodiment of a third aspect of the present invention is shown. The apparatus is adapted to prepare gene-transformed cells from a biological fluid. Specifically, the apparatus includes an operating unit 70 adapted to receive a processing chamber 100 and to orient the processing chamber about a rotation axis 71 of the operating unit. Specifically, the apparatus 300 is adapted to orient the central axis 101 of the processing chamber 100 vertically and / or horizontally or obliquely (the surface line 16 of the first body segment 1 is horizontal). Figure 5aThe configuration of the device 300 is shown when the processing chamber 100 is vertically arranged (the central axis 101 is aligned in the vertical position). Figure 5b The configuration of the device 300 is shown in the case where the processing chamber 100 is arranged horizontally (the central axis 101 is aligned in a horizontal position).

[0069] The device 300 allows for the execution of an advantageous method according to the fourth aspect of the invention because it enables the processing chamber 100 to be rotated from one position to another, wherein bringing the processing chamber to different positions corresponds to different method steps.

[0070] Advantageously, the device 300 includes a kit 200 according to the second aspect of the invention.

[0071] Figure 6 The flowchart steps i to viii of an embodiment of a method for culturing and processing cells in a first embodiment of a processing chamber 100 are shown. Figures 1a to 1c A first embodiment of the processing chamber 100 is shown. For clarity in the drawings, reference numerals are only added to... Figure 6 The accompanying drawings are for steps i, iv, and vi.

[0072] The method includes the following steps: bringing the processing chamber 100 in such that the central axis 101 is vertically aligned to a position where the cells are subjected to centrifugation (a small arrow near and below the processing chamber indicates centrifugation (one arrow) and / or agitation (two arrows)); and bringing the processing chamber 100 in such that the central axis 101 is horizontally aligned to a position where the cell cluster is suspended and the cells are incubated.

[0073] In an advantageous embodiment, the step of bringing the processing chamber 100 into a position in which the central axis 101 is vertically aligned is performed before bringing the processing chamber 100 into a position in which the central axis 101 is horizontally aligned.

[0074] Figure 6 Step i illustrates a processing chamber 100 including cell suspension 102 (dashed area). Figure 6 In step ii, the cell suspension is centrifuged, causing cell clusters 103 to form laterally inner surfaces 15 near the first main body segment 1, respectively, between the first main body segment 1 and the second main body segment 2. Figure 6 As shown in step iv (particularly at the surface and at the angle or edge 17 formed between the first main body section 1 and the second main body section 2). When centrifugation slows down or stops, the supernatant 104 can be collected at the bottom of the processing chamber (meaning specifically in the third main body section 3). The supernatant 104 or any other fluid can be removed from the processing chamber 100 through the second opening 31. Figure 6 Step v).

[0075] Advantageously after the centrifugation step, such as Figure 6 As shown in step vi, the processing chamber 100 is brought into a position where the central axis 101 is horizontally aligned. Figure 6 As shown in step vii, any liquid or solution can be added to the processing chamber 100. The cell cluster 103 can now be suspended in the solution by stirring. Figure 6 Step viii). It can be as follows: Figure 6 Step viii further incubates the suspended cells to promote cell division. The advantage of the processing chamber 100 now comes with its increased volume and increased lateral inner surface 15, resulting from the innovative shape of the processing chamber 100.

[0076] Figure 7 Steps i to viii of another embodiment of the method for culturing and processing cells in the second embodiment of the processing chamber 100 are shown. Figures 2a to 2c A second embodiment of the processing chamber 100 is shown. For clarity in the drawings, reference numerals are only added to... Figure 7 The accompanying drawings are for steps i, iv, and vi.

[0077] The method includes the following steps: bringing the processing chamber 100 in such that the central axis 101 is vertically aligned with a position for centrifugation of the cells (arrows near and below the processing chamber indicate centrifugation and / or agitation of the processing chamber); and bringing the processing chamber 100 in such that the central axis 101 is horizontally aligned with a position for incubating the cells.

[0078] In an advantageous embodiment, the step of bringing the processing chamber 100 into a position in which the central axis 101 is vertically aligned is performed before bringing the processing chamber 100 into a position in which the central axis 101 is horizontally aligned.

[0079] Figure 7 Step i illustrates a processing chamber 100 including cell suspension 102 (dashed area). Figure 7 In step ii, the cell suspension is centrifuged, causing cell clusters 103 to form laterally inner surfaces 15 near the first main body segment 1, respectively, between the first main body segment 1 and the second main body segment 2. Figure 7 Step iv is shown (here, the lateral inner surface, in particular the surface and the angle or edge 17 formed between the first body section 1 and the second body section 2).

[0080] When centrifugation slows down or stops, the supernatant 104 can be collected at the bottom of the processing chamber (meaning specifically in the third main body section 3). The supernatant 104 or any other fluid can be removed from the processing chamber 100 through the second opening 31. Figure 7 v).

[0081] Advantageously after the centrifugation step, such as Figure 7 As shown in step vi, the processing chamber 100 is brought into a position where the central axis 101 is horizontally aligned. Figure 7 As shown in step vii, any liquid or solution can be added to the treatment chamber 100. The cell clusters can now be suspended in the solution by stirring. Figure 7 Step viii). It can be as follows: Figure 7 Step viii further incubates the suspended cells to promote cell division. The advantage of the processing chamber 100 now comes with its increased volume and increased inner shell surface 15, resulting from the innovative shape of the processing chamber 100.

[0082] Advantageously, such as Figure 6 and Figure 7 The embodiments of the method shown refer to methods that are advantageous for centrifuging and incubating suspended cells.

[0083] Figure 8 Steps of another embodiment of a method for advantageously treating adherent cells using the first embodiment of the treatment chamber 100 shown in FIG. 1 are illustrated. The same method for advantageously treating and growing adherent cells can also be performed using the second embodiment of the treatment chamber 100 shown in FIG. 2. The steps of the method using the second embodiment are not shown here, but can be understood from... Figure 8 And it is easy to apply.

[0084] Figure 8 Steps i to viii of the process for culturing and processing cells in the processing chamber 100 in a first embodiment are shown. Figures 1a to 1c A first embodiment of the processing chamber 100 is shown. For clarity in the drawings, reference numerals are only added to... Figure 8 The accompanying drawings are for steps i, ii, iii, iv, and vi.

[0085] The method includes the following steps: bringing the processing chamber 100 in such a way that the central axis 101 is vertically aligned to the location for collecting liquid in the third main section. Figure 8 Steps iv and v); and bringing the processing chamber 100 in such that the surface line 16 of the first body section 1 is horizontally aligned with the position for incubating the cells ( Figure 8 Steps i to iii and Figure 8 Steps vi to vii).

[0086] In an advantageous embodiment, the step of bringing the processing chamber 100 into a position where the surface line 16 of the first main body section 1 is horizontally aligned on the horizontal plane 110 (horizontal position of the surface line of the first main body section) can be performed alternately with the step of bringing the processing chamber 100 into a position where the central axis 101 is vertically aligned (vertical position).

[0087] like Figure 8 As shown in step ii, a suspension 102 of adherent cells in the growth medium is added to the treatment chamber 100.

[0088] like Figure 8 As shown in step iii, when the processing chamber 100 rotates, the cells will form an adherent layer 105 on the lateral inner surface of the first body segment 1. Since the processing chamber 100 is aligned such that the surface line 16 of the first body segment 1 is horizontally aligned along the horizontal plane 110, the cells can advantageously adhere to the lateral inner surface 15 of the first body segment 1.

[0089] Furthermore, particularly when the processing chamber is positioned at the horizontal level of the surface line of the first main section ( Figure 8 (Step iii) can be used to add growth medium 106 to treatment chamber 100.

[0090] In the vertical position of the processing chamber 100 (meaning the processing chamber 100 is brought into a position where the central axis 101 is vertically aligned), the growth medium 106 can be collected. Figure 8 Step v) removes the growth medium again through the first opening 13 at the first end 10 of the treatment chamber 100 or through the second opening 31 at the second end 30 of the treatment chamber 100. Figure 8 Step v), while the adherent cell layer 105 remains on the lateral inner surface 15 of the first main body section 1.

[0091] Advantageously, after growth, trypsinizing agent 107 can be added to the treatment chamber through a first opening 13 at the first end of the treatment chamber and / or through a second opening 31 at the second end of the treatment chamber. Figure 8 vi). In order to add trypsinizing agent 107, in particular, the treatment chamber 100 can be positioned at the level of the surface line of the first main body section (vi). Figure 8 Steps vi and vii). The trypsinizing agent 107 can release cells attached to the lateral inner surface 15 of the first host segment 1 by cleaving peptide bonds. Thus, the adherent cells or adherent cell layer 105 are suspended in trypsin solution or in trypsin agent, respectively. Figure 8 (step vii), and can be removed from the processing chamber 100 for further use, for example by bringing the processing chamber into an upright position and releasing cells through a first opening 13 at the first end 10 of the processing chamber 100 or through a second opening 31 at the second end 30 of the processing chamber 100. Figure 6 Step viii), or by following Figure 6 The steps shown in steps i to v wash the cells out of the processing chamber using trypsin.

[0092] Reference Table

[0093] 1 First main section

[0094] 10. First end of the processing chamber

[0095] 11 First base side

[0096] 12 Second base side

[0097] 13 The first opening of the processing chamber

[0098] 15 Lateral inner shell surface of the first main section

[0099] 16 Surface line of the first main section

[0100] 17 Edge

[0101] 100 Centrifuge Processing Chamber

[0102] 101 Central Axis

[0103] 102 Cell suspensions

[0104] 103 cell clusters

[0105] 104 Supernatant

[0106] 105 (adherent) cell layers

[0107] 106 (cell) growth medium

[0108] 107 Trypsinizing agents

[0109] 110 horizontal plane

[0110] 2 Second main section

[0111] 200 kits

[0112] 3. Third main section

[0113] 30 The second end of the processing chamber

[0114] 300 device

[0115] 31 The second opening of the processing chamber

[0116] 40 First Flow Stator

[0117] 41 Rotary seal

[0118] 42 bearing

[0119] 43 Filters

[0120] 50 Second Circulation Stator

[0121] 60 Pipeline Group

[0122] 61. Reactant or product bag

[0123] 70 operating units

[0124] 71 Rotation axis of the operating unit

Claims

1. A processing chamber (100) concentrically formed around a central axis (101), the processing chamber comprising: - A first main body section (1) in the shape of a truncated cone, the first main body section including a first base side (11) and a second base side (12), wherein the first base side (11) has a smaller diameter than the second base side, and forms the first end (10) of the processing chamber (100) in the axial direction of the central axis (101). The second base side (12) leads to the second main body section (2), wherein, - The second main body segment (2) extends along the central axis (101), and the first end of the second main body segment (2) points away from the first main body segment (1). The first end of the second main body section (2) extends into the third main body section (3). - The third main body section (3) extends along the central axis (101), and the first end of the third main body section (3) points away from the second main body section (2), wherein the first end of the third main body section (3) tapers to the second end (30) of the processing chamber (100).

2. The processing chamber (100) according to claim 1, wherein, The slope (a) of the second main body section (2) relative to the central axis is greater than the slope (b) of the third main body section (3) relative to the central axis.

3. The processing chamber (100) according to any one of the preceding claims, wherein, The second main body segment is a truncated second main body segment (2), specifically including a first base segment of the second main body segment (2) closer to the first main body segment (1), the diameter of the first base segment being larger than the diameter of the second base segment of the second main body segment (2).

4. The processing chamber (100) according to claim 3, wherein, The height h of the truncated second main body section (2) f for h f ≤10mm, specifically h f ≤5mm.

5. The processing chamber (100) according to claim 1 or 2, wherein, The surface line of the second main body section (2) follows a generally elliptical or circular outline in the surface line that enters the third main body section (3).

6. The processing chamber (100) according to any one of the preceding claims, wherein, The minimum angle α between the tilt height of the second main body section (2) and the second base side (12) of the first main body section (1) is α≤20°, particularly α≤10°, particularly α≤5°.

7. The processing chamber (100) according to any one of the preceding claims, the processing chamber comprising a first opening for flow exchange at the first end (10) and / or a second opening for flow exchange at the second end (30).

8. The processing chamber (100) according to any one of the preceding claims, the processing chamber comprising a first flow stator (14) at the first end (10) and / or a second flow stator (31) at the second end (30).

9. The processing chamber (100) according to any one of the preceding claims, wherein, The first main body section (15) has at least an inner surface coated with a bioactive coating.

10. A kit (200) comprising: The processing chamber (100) according to any one of the preceding claims is connected to the first flow stator (40) and / or the second flow stator (50). A pipe assembly (60) capable of connecting to the first flow stator (40) and / or the second flow stator (50), and adapted to connect with at least one peristaltic pump head and at least one pinch valve in the intended use of the kit (200). At least one reactant or product bag (61) is adapted to be connected to the pipeline assembly (60).

11. An apparatus (300) suitable for preparing gene-transformed cells from a biological fluid, the apparatus particularly comprising an operating unit (70) adapted to receive a processing chamber (100) according to any one of claims 1 to 9, and adapted to orient the processing chamber about a rotation axis (71) of the operating unit (70), particularly adapted to orient the central axis (101) of the processing chamber (100) vertically and / or horizontally. Specifically, the device includes the kit (200) according to claim 10.

12. A method for culturing and treating cells in a processing chamber according to any one of claims 1 to 9, the method comprising the steps of: Bring the processing chamber (100) into a position where the central axis (101) is vertically aligned to subject the cells to centrifugation, and / or Bring the processing chamber (100) in such a way that the central axis (101) is horizontally aligned with the position for incubating cells, and / or Bring the processing chamber in such that the surface line of the first main body section (16) is horizontally aligned with the position for incubating the cells.

13. The method of claim 12, wherein the method is used to prepare gene-transformed cells from a biological fluid using the processing chamber (100) and the apparatus (300) of claim 11, the method comprising: Rotate the operating unit (70) to a position where the central axis (101) of the processing chamber (100) is in a vertical position, and / or Rotate the operating unit (70) to a position where the central axis (101) of the processing chamber (100) is horizontal, and / or Rotate the operating unit (70) to a position where the surface line of the first body section (16) is horizontally aligned to incubate the cells.

14. The method according to claim 12 or 13, wherein, The liquid or solution in the processing chamber (100) is exchanged via the first opening (13) and / or the second opening (31).

15. Use of the processing chamber (100) according to claims 1 to 9 in a method for preparing gene-transformed cells from biological fluids, wherein the method is preferably the method defined in any one of claims 12 to 14.

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