High-Flow Filter and Method of Use

Abstract

A filter is disclosed. The filter is a flexible container that defines an inlet port toward an inlet end of the container, an outlet port toward an outlet end of the container, and has one or more walls connecting the inlet end and the outlet end, a container, and a first filter unit and a second filter unit disposed spaced apart within the container, wherein the first filter unit defines an inlet chamber coupled to the container and in fluid communication with the inlet port, the second filter unit defines an outlet chamber coupled to the container and in fluid communication with the outlet port, the first filter unit and the second filter unit define an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit is configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit is configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the first filter unit and the second filter unit, and the inlet chamber includes a first trough with respect to a first peak, the intermediate chamber includes a second trough with respect to a second peak, and the outlet port and the second filter unit are configured to be spaced apart from each other when fluid flows into the outlet chamber through the second filter unit.

Description

Technical Field

[0001] Priority Application This application claims priority and benefit to U.S. Provisional Application No. 63 / 365,393, filed May 26, 2022, and U.S. Provisional Application No. 63 / 482,768, filed Feb. 1, 2023, the entire contents of each of which are incorporated herein by reference for all purposes.

[0002] The present disclosure relates to an apparatus for preparing a cell composition with reduced particles and / or cell aggregates, and a method of using the apparatus. Some embodiments of the present disclosure relate to a filtration apparatus configured to maintain a relatively high flow rate of a filtered cell composition exiting the filtration apparatus.

Background Art

[0003] Several cell therapy products for regenerative therapy or immunotherapy applications have advanced to clinical evaluation and market approval. The manufacturing processes for such products typically involve culturing cells in the presence of non-autologous serum and harvesting cells by trypsin digestion.

[0004] At several stages during the manufacturing process, the cells are exposed to external substances. At these stages, there is a risk that the cells can be contaminated with one or more particles such as cotton fibers, cellulose, salt crystals, rubber, plastic, glass, etc. The presence of such particles in the final product is potentially harmful to the cells and / or the individuals receiving the resulting cell therapy. Generally, filtration is required to remove particles before the final composition to be used in therapy is provided. However, filtering cell compositions is not straightforward considering one or more factors such as the complex nature of cell culture media and the tendency of cells to aggregate, especially in large-scale cell cultures. Clearly, there are unmet needs in the art for techniques for preparing cell compositions, particularly in the field of therapeutic cell compositions.

[0005] Any discussion of documents, acts, materials, devices, articles, etc. included in this specification shall not be construed as an admission that any or all of their contents form part of the prior art base as of the priority date of each of the appended "claims" or were common general knowledge in the field related to this disclosure.

[0006] Throughout this specification, words such as "comprise", "comprises" or "comprising" shall be understood not to exclude the inclusion of a particular element, component or step, or group of elements, components or steps, but not to exclude any other element, component or step, or group of elements, components or steps.

SUMMARY OF THE INVENTION

MEANS FOR SOLVING THE PROBLEM

[0007] Some embodiments relate to a filter, the filter comprising a flexible container defining an inlet port towards an inlet end of the container and an outlet port towards an outlet end of the container, the container having one or more walls connecting the inlet end and the outlet end, a first filter unit and a second filter unit spaced apart within the container, the first filter unit being coupled to the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the inlet chamber comprising a first trough with respect to a first peak, the intermediate chamber comprising a second trough with respect to a second peak, and the outlet port and the second filter unit being configured to be spaced apart from each other when fluid flows into the outlet chamber through the second filter unit.

[0008] Some embodiments relate to a filter, the filter being a container that defines an inlet port toward an inlet end of the container, an outlet port toward an outlet end of the container, and having a flexible front wall and a rear wall connecting the inlet end and the outlet end, a container, and a first filter unit and a second filter unit spaced apart within the container, the first filter unit being coupled to the front wall and the rear wall of the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the front wall and the rear wall of the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the first filter unit being coupled to the front wall along a first front seam, the second filter unit being coupled to the front wall along a second front seam, the first filter unit being coupled to the rear wall at a first back seam, the second filter unit being coupled to the rear wall at a second back seam, the distance between the first front seam and the second front seam being less than the distance between the first back seam and the second back seam, and the outlet port and the second filter unit being configured to be spaced apart from each other when fluid flows into the outlet chamber through the second filter unit.

[0009] The container can be a flexible bag. The walls can be flexible. The distance between the first front face and the second front face can be 1 inch or less. The second back seam can be angled away from the second filter unit and the outlet port relative to each other. The front wall can be configured to bulge in a direction away from the second filter unit.

[0010] The outlet port and the second filter unit may be configured to be separated by a spacer system. The spacer system may be configured to separate the outlet port and the second filter unit, and may also be configured to separate at least one of (i) the front wall and the first filter unit, (ii) the rear wall and the first filter unit, and (iii) the rear wall and the second filter unit.

[0011] The spacer system may include a separator. The spacer system may include clips. The spacer system may include a magnet system. The spacer system may include braces.

[0012] The first filter unit may be connected to the front wall of the container at an acute angle to define an inlet chamber trough. The first filter unit may be connected to the rear wall of the container at an obtuse angle. The first filter unit may include a mesh defining holes having an average pore size of 130 μm to 170 μm. The holes of the first filter unit may have an average pore size of 150 μm. The first filter unit may be substantially flat.

[0013] The second filter unit may be connected to the rear wall of the container at an acute angle to define an intermediate chamber trough. The second filter unit may be connected to the front wall of the container at an acute angle. The second filter unit may include a mesh defining holes having an average pore size of 20 μm to 60 μm. The holes of the second filter unit may have an average pore size of 40 μm. The second filter unit may be substantially flat.

[0014] The inlet chamber may have a volume that is about 0.9 to 3.2 times the surface area of the first filter unit. The intermediate chamber may have (i) a volume that is about 1.9 to 4.0 times the surface area of the first filter unit and (ii) a volume that is about 2.7 to 5.6 times the surface area of the second filter unit. The outlet chamber may have a volume that is about 1.6 to 4.5 times the surface area of the second filter unit. The volume ratio of the inlet chamber, the intermediate chamber, and the outlet chamber is about (i) 1:1:1, or (ii) 1:2:1, or (iii) 1:2:2, or (iv) 2:2:1, or (v) 1:3:2.

[0015] When viewed parallel to the direction of fluid flow from the inlet port to the outlet port, at least one of the chambers may have a substantially triangular or substantially trapezoidal cross-section.

[0016] Some embodiments relate to a filter, the filter being a flexible bag that defines an inlet port toward the inlet end of the bag, an outlet port toward the outlet end of the bag, and having opposing front and rear walls connecting the inlet end and the outlet end, the bag, a first filter unit and a second filter unit spaced apart within the bag, the first filter unit defining an inlet chamber coupled to the bag and in fluid communication with the inlet port, the second filter unit defining an outlet chamber coupled to the bag and in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, and the outlet port and the second filter unit being configured to be spaced apart from each other when fluid flows into the outlet chamber through the second filter unit.

[0017] The first filter unit and the second filter unit may not be in parallel.

[0018] Some embodiments relate to a kit for filtering cells, the kit comprising the filter described above, an inlet conduit comprising a first inlet tube and a second inlet tube fluidly connected to a manifold, and a filling tube in fluid communication with the manifold and adapted to receive fluid from at least one of the first inlet tube and the second inlet tube, and an inlet coupling connecting the filling tube to an inlet port of the filter and adapted to enable fluid communication with an inlet chamber, an outlet conduit comprising a drain tube and an outlet coupling connecting the drain tube to an outlet port of the filter and adapted to enable fluid communication between an outlet chamber and the drain tube.

[0019] The inlet tube and the filling tube may form a Y shape or a T shape. The inlet tube may have an inner diameter of 1 / 8 inch and an outlet diameter of 1 / 4 inch. The filling tube may have an inner diameter of 1 / 8 inch and an outlet diameter of 1 / 4 inch. The drain tube may have an inner diameter of 1 / 8 inch and an outlet diameter of 1 / 4 inch. The kit may further comprise an outlet valve or clamp configured to control the flow rate of fluid passing through the drain tube.

[0020] One or more or all of the container, bag, filter units 120, 130, or tubes may be made of DMSO (dimethyl sulfoxide)-compatible plastic, preferably plastic comprising one or more of polyethylene terephthalate (PET), high density polyethylene (HDPE), low density polyethylene (LDPE), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), thermoplastic elastomer (TPE), and polypropylene (PP).

[0021] Some embodiments relate to a filter, the filter comprising a container defining an inlet port towards an inlet end of the container and an outlet port towards an outlet end of the container, the container having one or more walls connecting the inlet end and the outlet end, a first filter unit and a second filter unit spaced apart within the container, the first filter unit being coupled to the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the inlet chamber comprising a first trough relative to a first peak, and the intermediate chamber comprising a second trough relative to a second peak.

[0022] Some embodiments relate to a filter, the filter being a container that defines an inlet port towards an inlet end of the container and an outlet port towards an outlet end of the container, and having opposing front and rear walls connecting the inlet end and the outlet end, a first filter unit and a second filter unit spaced apart within the container, the first filter unit being coupled to the front and rear walls of the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the front and rear walls of the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the first filter unit being coupled to the front wall along a first front seam, the second filter unit being coupled to the front wall along a second front seam, the first filter unit being coupled to the rear wall at a first back seam, the second filter unit being coupled to the rear wall at a second back seam, and a distance between the first front seam and the second front seam being smaller than a distance between the first back seam and the second back seam.

[0023] Some embodiments relate to a filter, the filter being a flexible bag defining an inlet port towards an inlet end of the bag, an outlet port towards an outlet end of the bag, and having opposing front and rear walls connecting the inlet end and the outlet end, a bag, a first filter unit and a second filter unit disposed spaced apart within the bag, the first filter unit defining an inlet chamber coupled to the bag and in fluid communication with the inlet port, the second filter unit defining an outlet chamber coupled to the bag and in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, and the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber.

[0024] The filter or kit described above can be used for filtering a stem cell culture medium.

[0025] Some embodiments relate to a method of filtering a stem cell culture medium, the method including using the filter or kit described above.

[0026] Some embodiments relate to a method of purifying a cell composition, the method including passing cultured cells through a double screen mesh filter to thereby reduce visible particles and / or cell aggregates, the double screen mesh filter including a first filter screen having an average pore size of 130 μm to 170 μm and a second filter screen having an average pore size of 20 μm to 60 μm.

[0027] The cultured cells can be provided in a serum-free cell culture medium. The cells can be cultured and proliferated. In one example, the cells are mesenchymal progenitor cells or stem cells (MLPSC).

[0028] After passing the cells through the double screen mesh filter, the recovery rate of viable cell concentration is (i) 60% to 100%, or (ii) 70% to 90%.

[0029] The purified cell composition may exhibit a D90 of less than 150 μm, preferably less than 100 μm, more preferably less than 50 μm. The purified cell composition may be substantially free of visible particles.

[0030] Some embodiments relate to a filter, the filter being a flexible container defining an inlet port towards an inlet end of the container, defining an outlet port towards an outlet end of the container, and having one or more walls connecting the inlet end and the outlet end, the container, a first filter unit including a first filter mesh, and a second filter unit including a second filter mesh, the first filter unit and the second filter unit being spaced apart within the container, the first filter unit being coupled to the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the inlet chamber comprising a first trough for a first peak, the intermediate chamber comprising a second trough for a second peak, and at least one of the first filter mesh and the second filter mesh being configured to be spaced from the one or more walls when fluid is passing through the filter.

[0031] Some embodiments relate to a filter, the filter comprising a container defining an inlet port towards an inlet end of the container and an outlet port towards an outlet end of the container, the container having a flexible front wall and a rear wall connecting the inlet end and the outlet end, a first filter unit including a first filter mesh, and a second filter unit including a second filter mesh, wherein the first filter unit and the second filter unit are spaced apart within the container, the first filter unit being coupled to the front wall and the rear wall of the container to define an inlet chamber in fluid communication with the inlet port, the second filter unit being coupled to the front wall and the rear wall of the container to define an outlet chamber in fluid communication with the outlet port, the first filter unit and the second filter unit defining an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit being configured to filter fluid flowing from the inlet chamber to the intermediate chamber, the second filter unit being configured to filter fluid flowing from the intermediate chamber to the outlet chamber, the first filter unit being coupled to the front wall along a first front seam, the second filter unit being coupled to the front wall along a second front seam, the first filter unit being coupled to the rear wall at a first back seam, the second filter unit being coupled to the rear wall at a second back seam, the distance between the first front seam and the second front seam being smaller than the distance between the first back seam and the second back seam, the outlet port and the second filter unit being configured to be spaced apart from each other when fluid flows into the outlet chamber through the second filter unit, and at least one of the first filter mesh and the second filter mesh being configured to be spaced apart from the front wall or the rear wall when fluid passes through the filter.

Brief Description of the Drawings

[0032]

Figure 1

Figure 2A

Figure 2B

Figure 2C

Figure 2D

Figure 3A

Figure 3B

Figure 4

Figure 5

Figure 6

DETAILED DESCRIPTION OF THE INVENTION

[0033] The present disclosure relates to an apparatus for preparing a cell composition with reduced particles and / or cell aggregates, and a method of using the apparatus.

[0034] In particular, some embodiments of the present disclosure relate to a filtration device suitable for filtering a cell composition or a fluid material such as a cell culture medium or a resuspension medium containing the same. As an example, the cell composition includes mesenchymal progenitor cells or stem cells. Some embodiments of the present disclosure relate to a filtration device configured to maintain a relatively high flow rate of the filtered cell composition exiting the filtration device. The device may be referred to as a "double screen mesh filter". The double screen mesh filter includes a first filter unit and a second filter unit. In some embodiments, the filter unit includes a screen or sheet made of a mesh material (thus referred to as a "double screen mesh filter"). The double screen mesh filter tends to be less prone to clogging compared to other filters when filtering a viscous fluid. Other embodiments of the filter may include pleated filters or depth filters.

[0035] In some embodiments, the filtration device may have sufficient flexibility to be deformable to fit into narrow or irregularly shaped locations. In certain applications, using a flexible filtration device may be particularly advantageous, especially when compared to a more rigid structure, as the flexible filtration device can be easily positioned around the more rigid components of an overall cell culture and / or cell purification system, including rigid structures such as stands, supports, pumps, etc. The filtration device with this flexibility may include flexible portions. In some embodiments, the filtration device is a flexible bag similar to a saline bag.

[0036] The filter unit is arranged such that during operation, the fluid passing through the device passes through both the first filter unit and the second filter unit and is then discharged outside the device. The filter unit may include a mesh with a selectable average pore size according to the size of the particles to be filtered from the flowing material. For example, a filter unit including a mesh with an average pore size of 150 μm theoretically prevents particles with a diameter exceeding 150 μm from passing through the filter unit. When the fluid passes through a series of filters, the pore size of the subsequent filters may decrease to achieve stepwise filtration. This can help reduce clogging of the filters. For example, the first filter unit may have an average pore size of 150 μm, and the second filter unit may have an average pore size of 40 μm.

[0037] The inventors have confirmed that in some situations, one or both of the mesh filters can bend or wrinkle. The mesh filter(s) may bend or wrinkle when the fluid passes through the mesh filter by gravity. In some situations, fluid may accumulate on the inlet side of the filter unit when the fluid enters the filtration device faster than it passes through each of the filter units within the filtration device.

[0038] The mesh filter(s) will not readily bend or wrinkle, but when a certain amount of fluid accumulates, the mesh filter(s) may bend under the weight of the accumulated fluid. The bending or wrinkling of the mesh filter(s) may cause a part of the mesh filter(s) to approach or contact the wall surface or inner surface of the filtration device, and as a result, may inhibit the flow of fluid passing through the corresponding part of the mesh filter (by restricting or blocking it). Such inhibition of fluid flow is likely to occur in flexible or bag-shaped filtration devices. This is because such filtration devices have low (or no) rigidity, making it easy for the filter(s) and the wall to approach each other.

[0039] The inventors have developed embodiments of devices and techniques for reducing or preventing relative movement between the filter(s) and the wall of the filtration device.

[0040] The inventors have developed embodiments of devices and techniques for reducing or preventing relative movement between the filter(s) and the wall of the filtration device while maintaining the flexibility of the filtration device.

[0041] Advantageously, by providing the filtration device of the described embodiments, it is possible to prevent or avoid the collapse of the filtration device (such as a flexible bag) due to gravity. In some embodiments, the filtration device of the described embodiments can reduce the restriction of fluid flow by preventing or guarding against the contact of the filter(s) (such as a flexible bag) with the wall of the filtration device.

[0042] According to the disclosed embodiments, the flow of fluid passing through the filtration device can become more uniform. According to the disclosed embodiments, the flow rate of fluid passing through the filtration device can be consistently higher compared to other filtration devices that do not use the disclosed embodiments. The disclosed embodiments will be described in more detail below, with particular reference to FIGS. 2A - 2D and FIGS. 3A, 3B.

[0043] Filtration device FIG. 1 shows an embodiment of a filtration device 100 for filtering fluids such as cell compositions. The filtration device 100 may also be referred to as a filter. The filter 100 includes a container 102 and a first filter unit 120 (only a part is shown) and a second filter unit 130 (only a part is shown) disposed within the container 102. The container 102 is adapted to receive the fluid, and the first filter unit 120 and the second filter unit 130 are configured to filter the fluid passing through the container 102. Each of the filter units 120, 130 may include a filter mesh having pores sized such that a particular size / type of substance can pass through the filter units 120, 130. The filter mesh of the first filter unit 120 may have pores of the same or different sizes as the filter mesh of the second filter unit 130.

[0044] The pores are defined by a structure that defines the pores, such as the body of the mesh. The mesh body and the container 102 may be made of a material that is compatible (e.g., resistant) with dimethyl sulfoxide (DMSO). Examples of DMSO-compatible materials include DMSO-compatible polymers. In some embodiments, the filter units 120, 130 comply with USP 788, which is a particulate matter test that quantifies the number and size of non-visible particles in a parenteral drug. The USP 788 test involves using a light-blocking particle counter and counting the particles on the filter by microscopy.

[0045] In some embodiments, the first filter unit 120 has an average pore size between 130 μm and 170 μm, or between 140 and 160 μm. In some embodiments, the first filter unit 120 has an average pore size of about 150 μm.

[0046] In some embodiments, the second filter unit 130 has an average pore size between 20 μm and 60 μm, or between 30 and 50 μm. In some embodiments, the second filter unit 130 has an average pore size of about 40 μm.

[0047] In some embodiments, the container 102 is a hollow body. The body may have a regular shape such as substantially square (cubic) or rectangular (cuboid) like a carton. The container 102 may be rigid enough to support its own weight or may be flexible enough to be placed in a narrow or irregularly shaped location. The container 102 may include a combination of rigid and flexible parts. In some embodiments, the container 102 is a flexible bag similar to a saline bag.

[0048] The container 102 has an inlet end 104 and an outlet end 106. The container 102 defines an inlet port 108 on the inlet end 104 side and an outlet port 110 on the outlet end 106 side. In some embodiments, the inlet port 108 and the outlet port 110 are arranged substantially opposite to each other such that when the filtration device 100 is in use, fluid flows from the inlet port 108 towards the outlet port 110 by gravity. In some embodiments, the container 102 includes a flange 112. The flange 112 may be disposed at the inlet end 104. The flange 112 may define an opening 114 or a connector through which the container 102 can be hung on a hook (not shown). The opening 114 enables the container 102 to be suspended or supported in an upright position, so that fluid can flow from the inlet port 108 towards the outlet port 110 by gravity while the filtration device 100 is in use.

[0049] Figures 2A - 2D are side views of the filtration device 100. The container 102 includes one or more walls 200 that connect an inlet end 104 and an outlet end 106. The one or more walls 200 may include a front wall 202 and a rear wall 204. The one or more walls 200 may further include side walls, sides, or side ends (not shown) that connect the front wall 202 and the rear wall 204 to define the body of the container 102. In some embodiments, the front wall 202 and the rear wall 204 are opposite (oppositely arranged). Some embodiments of the container 102 may further include an upper connection portion 210 and a lower connection portion 212. The upper connection portion 210 and the lower connection portion 212 may be seams that connect the front wall 202 and the rear wall 204. The seams 210 and the flange 112 may be connected to each other. For example, the flange 112 may extend from the seam 210. In some embodiments, the inlet end 104, the outlet end 106, and the side ends include at least one seam connecting the one or more walls 200 or edges welded / bonded in a sterilized state.

[0050] The first filter unit 120 and the second filter unit 130 are spaced apart within the container 102 and are connected to the wall 200 of the container 102. By spacing the first filter unit 120 and the second filter unit 130 apart, more fluid can pass through the first filter unit 120 and then through the second filter unit 130. The spaced - apart filter units 120, 130 can reduce the possibility of filter clogging, for example, by the structure where the first filter unit 120 and the second filter unit 130 contact each other and the holes of one filter unit block the holes of the other filter unit. The filter units 120, 130 may also stick to each other when the fluid to be filtered is sticky or particularly viscous.

[0051] When the filtration device 100 is suspended so that fluid passes through the filtration device 100 by gravity, the first filter unit 120 and the second filter unit 130 can usually be in a relatively taut state without bending in any direction of the wall 200. The taut first filter unit 120 and second filter unit 130 are labeled 120-1 and 130-1, respectively.

[0052] As described above, when fluid passes through the first filter unit 120 and heads towards the outlet port 110, due to the weight of the fluid on the first filter unit 120, at least a part of the first filter unit 120 can bend and move in a direction away from the inlet port 108. The bent state of the first filter unit 120 is labeled 120-2. In the bent state 120-2, at least a part of the first filter unit 120 approaches or contacts the wall 200 of the container 102, and as a result, the flow of fluid passing through that part of the first filter unit 120 may be obstructed or blocked. For example, in the bent state 120-2, at least a part of the first filter unit 120 can contact the rear wall 204. Thereby, the surface area of the first filter unit 120 available for filtering fluid can be reduced.

[0053] Similarly, when the fluid passes through the second filter unit 130 and towards the outlet port 110, due to the weight of the fluid on the second filter unit 130, at least a part of the second filter unit 130 can deflect and move away from the inlet end 104. The deflected state of the second filter unit 130 is labeled as 130-2. In the deflected state 130-2, at least a part of the second filter unit 130 approaches or contacts the wall 200 of the container 102, whereby the flow of the fluid passing through that part of the second filter unit 130 may be obstructed or blocked. For example, in the deflected state 130-2, at least a part of the second filter unit 130 can contact the front wall 202. In some situations, at least a part of the second filter unit 130 may contact the outlet port 110. As a result, the surface area of the second filter unit 130 available for filtering the fluid can be reduced.

[0054] The above-described reduction in the filtration surface area of the first filter unit 120 and the second filter unit 130 can substantially affect the flow rate of the filtered fluid passing through the outlet port 110.

[0055] The inventors have identified that in some embodiments, the flow rate of the filtered fluid passing through the outlet port 110 may be more significantly affected by the movement of the second filter unit 130 towards the wall 202 than by the movement of the first filter unit 120 towards the wall 204. The inventors believe that this may be because the second filter unit 130 may have a smaller pore size on average than the first filter unit 120.

[0056] The inventors believe that when the filtration device 100 is a saline bag or similar, the shape of the bag may affect the flow of fluid passing through the second filter unit 130. This is shown, for example, in FIG. 2A, which depicts a container 102 with end portions 104, 106 that are narrower compared to the central portion therebetween. Thus, when the second filter 130 is in the bowed state 130-2, the distance until the second filter unit 130 contacts the wall 202 of the container 102 is shortened. In contrast, since the central portion of the container 102 in FIG. 2A is wide, when the first filter unit 120 is in the bowed state 120-2, the distance until the first filter unit 120 contacts the wall 204 of the container 102 is longer.

[0057] In some embodiments, the outlet port 110 and the second filter unit 130 are configured to be spaced apart from each other. When fluid passes through the second filter unit and flows into the outlet chamber, the outlet port 110 and the second filter unit 130 can be configured to be spaced apart from each other. In embodiments where the container 102 is flexible, such as a saline bag, the distance between the outlet port 110 and the second filter unit 130 may vary, and in some configurations, the outlet port 110 and the second filter unit 130 may contact each other, while in other configurations, the outlet port 110 and the second filter unit 130 are arranged to be spaced apart.

[0058] Referring to FIGS. 2A-2D, the filtration device 100 may include a spacer system 220. The spacer system 220 is configured to space apart the outlet port 110 and the second filter unit 130. The spacer system 220 can cause the outlet port 110 and the second filter unit 130 to contact and separate by allowing the container 102 to vary between a contracted state and an expanded state.

[0059] By spacing the second filter unit 130 away from the front wall 202, the spacer system 220 keeps the holes of the second filter unit 130 unblocked or in a relatively clear state, allowing fluid to continue passing through the second filter unit 130 and into the outlet chamber 320.

[0060] The spacer system 220 can enable the flow rate of the fluid passing through the second filter unit 130 to be maintained constant (or with a minimal decrease), regardless of whether the second filter unit 130 is in the stretched state 130-1 or the deflected state 130-2.

[0061] FIG. 2A shows an embodiment of the spacer system 220 in the form of a separator 222. The separator 222 includes a separator body 224. The separator 222 is configured to be connected to the outlet port 110 inside the container 102. Thereby, the separator 222 and the outlet port 110 can sandwich the front wall 202.

[0062] When the second filter unit 130 is in the deflected state 130-2, the separator 222 inhibits the second filter unit 130 from moving towards the outlet port 110. The separator 222 can limit the surface area of the second filter unit 130 that can contact the front wall 202. FIG. 2A includes two inset views each showing an embodiment of the separator 222, labeled as inset view 1 and inset view 2. Inset view 1 is a side view showing how the separator 222 can limit the surface area of the second filter unit 130 that can contact the front wall 202 according to some embodiments. Inset view 2 is an end view (seen from the filter side) of the separator 222 according to some embodiments.

[0063] The separator body 224 can be disc-shaped, and the distance by which the second filter unit 130 is separated from the front wall 202 is set by its thickness. The separator 222 may have rigidity. The separator 222 may be made of the same material as the outlet port 110. The separator body 224 defines a plurality of openings through which fluid can flow to the outlet port 110.

[0064] The separator 222 may further include at least one ridge or fence 226 extending from the separator body 224, which can substantially increase the thickness of the separator 222 and widen the gap between the second filter unit 130 and the front wall 202. In some embodiments, the separator 222 includes at least two ridges or fences 226. The fence 226 and the separator body 224 may define an opening or channel 228 between adjacent fences 226. The channel 228 is configured such that fluid can pass through the channel, as indicated by the arrow FF in FIG. 2A (see Insertion Drawing 1 and Insertion Drawing 2). The fence 226 may be configured to direct the fluid passing through the channel 228 towards the outlet port 110. In some embodiments, some of the fences 226 may define an opening or passage, allowing fluid to flow between adjacent channels 228 towards the outlet port 110.

[0065] The fence 226 can be configured such that the outlet port 110 is not blocked by the second filter unit 130. When the second filter unit 130 is in the deflected state 130-2, the second filter unit 130 is configured to abut against the fence 226. The fence 226 is configured to support the second filter unit 130 and to space the second filter unit 130 apart from the separator body 224. Adjacent fences 226 can be spaced apart from each other such that the second filter unit 130 does not deflect significantly between the fences 226 and block the channel 228. Thereby, the outlet port 110 can be unblocked by the second filter unit 130. As a result, the fluid can pass through the channel 228 as indicated by the arrow FF in FIG. 2A (see inset 1 and inset 2).

[0066] In some embodiments, the fence 226 is configured to direct the fluid passing through the channel 228 towards the outlet port 110. For example, the fence 226 may extend radially from the outlet port 110, and the outlet port 110 may be disposed at the central portion of the separator body 224. In this configuration, all channels 228 can directly direct the fluid flow towards the outlet port 110.

[0067] FIG. 2B shows an embodiment of the spacer system 220 in the form of a clip 230. The clip 230 is configured to be connected to the container 102 and hold the outlet port 110 and the second filter 130 apart. In some embodiments, the clip 230 sandwiches the front wall 202 and the rear wall 204. Since the second filter 130 is connected to the front wall 202 and the rear wall 204, by holding the walls 202, 204 apart, tension is applied to the second filter 130, and the second filter 130 is held in a state pulled away from the outlet port 110.

[0068] FIG. 2C shows an embodiment of the spacer system 220 in the form of a magnet system 240. The magnet system 240 includes a first magnetic part 242 coupled to the front wall 202 and a second magnetic part 244 coupled to the rear wall 204. The magnetic parts 242, 244 are configured to be connected to the respective walls 202, 204. For example, it may be joined with an adhesive, or may be housed in a pouch created by joining / welding a layer of material to the walls 202, 204. The magnetic parts 242 and 244 are isolated from the fluid in the container 102 by the walls 202 and 204, so that the fluid in the container 102 does not come into contact with the magnetic parts 242, 244 (for example, to avoid contamination of the cell composition). The magnetic parts 242, 244 are preferably disposed near the outlet port 110.

[0069] In some embodiments, the magnetic parts 242, 244 are magnets respectively, and the polarities of the respective magnets are arranged such that the magnets 242, 244 repel each other, thereby pulling apart and holding the front wall 202 and the rear wall 204. When the filtration device 100 is in use, the magnets 242, 244 can be magnetically attracted to the adjacent metal structures 246, 248. The adjacent metal structures 246, 248 may be part of an exoskeleton (not shown) configured to support the filtration device 100. When the magnets 242, 244 engage the adjacent metal structures 246, 248, the magnet system 240 pulls apart and holds the front wall 202 and the rear wall 204.

[0070] FIG. 2D shows an embodiment of the spacer system 220 in the form of a brace 250. The brace 250 is configured to be applied to the second filter unit 130 and configured to support the mesh. The brace 250 may include a plurality of rigid members, which are arranged to reduce the deflection of the mesh of the second filter unit 130. The inventors have identified that the flow of fluid through the second filter unit 130 may be obstructed by a particular arrangement of the members. The rigid members of the brace 250 may be mainly disposed at or near the periphery of the second filter unit 130. The rigid members of the brace 250 may be thin wires.

[0071] In some embodiments, the brace 250 includes a convex portion such that the second filter unit 130 is formed in at least a partially convex shape as shown by the dashed line 250-1. The convex portion of the second filter unit 130 arches the second filter unit 130 away from the outlet port 110. This means that when fluid accumulates in the second filter unit 130, the weight of the fluid first acts to flatten the second filter unit 130, thereby reducing the likelihood of causing a deflection (concavity) of the second filter unit 130.

[0072] Embodiments of the spacer system 220 may be combined to operate in combination. For example, the separator 222 may be used in combination with the magnet system 240. The spacer system 220 can be used to space apart either the wall 200 and the filter units 120, 130, particularly in embodiments where the container 102 is flexible. The spacer system 220 can be configured to space apart the front wall 202 and the first filter unit 120. The spacer system 220 can be configured to space apart the front wall 202 and the second filter unit 130. The spacer system 220 can be configured to space apart the rear wall 204 and the first filter unit 120. The spacer system 220 can be configured to space apart the rear wall 204 and the second filter unit 130.

[0073] The foregoing embodiments of the spacer system 220 are described and shown in FIGS. 2A-2D as being applicable only to the second filter unit 130. However, the spacer system 220 can be similarly applied to the first filter unit 120 as well. For example, the separator 222 can be disposed on the rear wall 204 such that the rear wall 204 moves away from the first filter unit 120 when the first filter unit 120 is in the deflected state 120-2. In some embodiments, the spacer system 220 is applied to the first filter unit 120 instead of, or in addition to, being applied to the second filter unit 130.

[0074] As best shown in FIGS. 3A-3B, the first filter unit 120 and the second filter unit 130 are connected to the container 102 and divide the interior of the container 102 into a plurality of individual regions or chambers 300. The chambers 300 are configured to be in fluid communication with each other via the first filter unit 120 and the second filter unit 130. The container 102 may include an inlet chamber 310 and an outlet chamber 320, and an intermediate chamber 330 may be disposed between the inlet chamber 310 and the outlet chamber 320. The spaced-apart arrangement of the first filter unit 120 and the second filter unit 130 defines an intermediate chamber 330 between the inlet chamber 310 and the outlet chamber 320. The first filter unit 120 may be configured to filter fluid flowing from the inlet chamber 310 to the intermediate chamber 330, and the second filter unit 130 may be configured to filter fluid flowing from the intermediate chamber 330 to the outlet chamber 320.

[0075] In some embodiments, the first filter unit 120 is coupled to the container 102 and defines an inlet chamber 310. The inlet chamber 310 may extend from the inlet end 104 to the first filter unit 120 coupled to the container 102, and may be coupled to, for example, at least the front wall 202 and the rear wall 204. The first filter unit 120 may be coupled to the container 102 (such as the front wall 202) along a first front seam 340, or may be coupled to the container 102 (such as the rear wall 204) at a first back seam 350. The first front seam 340 and the first back seam 350 may extend to contact each other at the side ends 206, 208 in embodiments where the container 102 has a configuration similar to a saline bag.

[0076] The inlet chamber 310 is configured to be in fluid communication with the inlet port 108, and the fluid flowing into the inlet port 108 collects in the inlet chamber 310 and passes through the first filter unit 120. The first filter unit 120 is configured to filter a first substance (such as particles) contained in the fluid. For example, the first substance may be particles having a size larger than the pore size of the first filter unit 120. The first filter unit 120 may be configured to filter the first substance from the fluid flowing from the inlet chamber 310 to the intermediate chamber 330. The first substance remains on the surface of the first filter unit 120, and the rest of the fluid passes through.

[0077] When the first substance accumulates on the surface of the first filter unit 120, the pores of the first filter unit 120 are gradually blocked, thereby reducing the filtration efficiency. Accordingly, in some embodiments, the inlet chamber 310 may comprise a first trough 342. In some embodiments, the first trough 342 is a concave portion of the inlet chamber 310. The first trough 342 is disposed relative to the first peak 352. In some embodiments, the first peak 352 is a protrusion of the inlet chamber 310 and may optionally be pointed. In some embodiments, the first peak 352 is defined by the first filter unit 120. The first trough 342 may also be defined relative to the first peak 352 of an adjacent intermediate chamber 330 (located on the opposite side of the first filter unit 120). In use, the first trough 342 is the lowermost portion of the inlet chamber 310, and the first peak 352 is part of the adjacent intermediate chamber 330 and is disposed at a position higher than the first trough 342 (i.e., closer to the inlet end 104). In some embodiments, the inlet chamber 310 may be substantially wedge-shaped. For example, the inlet chamber 310 may be shaped such that it gradually narrows from the inlet end 104 or near it towards the first trough 342.

[0078] The first filter unit 120 may define the first trough 342 by being angled and connected to at least one of the container walls 200, and may define the first peak 352 by being angled and connected to another container wall 200. For example, the first trough 342 may be defined by the first filter unit 120 and the front wall 202. The angle (measured within the inlet chamber 310) defining the first trough 342 may be an acute angle. The first trough 342 may sometimes be referred to as the inlet chamber trough. The first peak 352 may be defined by the first filter unit 120 and the rear wall 204. The angle (measured within the intermediate chamber 330) defining the first peak 352 may be an obtuse angle. When such an embodiment is used, the first filter unit 120 is inclined inside the container 102.

[0079] In use, the first filter unit 120 can be configured to direct the first substance into the first trough 342. The inclination of the first filter unit 120 means that particles that do not pass through the filter unit 120 move along the filter mesh into the trough 342 of the inlet chamber 310. For example, since the trough 342 is located at the bottom of the inlet chamber 310, these particles can be urged by gravity to settle in the trough 342 rather than on the filter surface. In some embodiments, the fluid in the inlet chamber 310 can wash the first substance along the surface of the first filter unit 120 and direct it to the first trough 342.

[0080] By inducing the first substance to accumulate in the first trough 342, more portions of the filter unit surface can be kept clear or unclogged, thus improving the filtration efficiency and / or shortening the time until the filter unit 120 (or the filtration device 100) needs to be cleaned or replaced. In contrast, if a horizontally arranged filter unit is used, these "discharged" particles (the first substance) can adhere to and accumulate in the holes of the horizontally arranged filter unit, potentially obstructing the fluid flow through it. Thus, by directing the "excluded" particles into the trough 342, the inclined filter unit 120 reduces the risk of pore blockage and allows more fluid to pass through the filter unit 120.

[0081] The fluid is configured to pass through the first filter unit 120 and enter the next chamber, which in some embodiments is the intermediate chamber 330. The upper part of the intermediate chamber 330 is defined by the first filter unit 120. In some embodiments, the second filter unit 130 is coupled to the container 102 (and is disposed spaced apart from the first filter unit 120) and defines the bottom of the intermediate chamber 330. In some embodiments, when the second filter unit 130 is coupled to the container 102, one side of the second filter unit 130 defines the bottom of the intermediate chamber 330 and the other side of the second filter unit 130 defines the outlet chamber 320.

[0082] In some embodiments, the intermediate chamber 330 includes a second trough 372. In some embodiments, the second trough 372 is a concave portion of the intermediate chamber 330. The second trough 372 is disposed relative to the second peak 362. In some embodiments, the second peak 362 is a protrusion of the intermediate chamber 330 and may optionally be pointed. In some embodiments, the second peak 362 is defined by the second filter unit 130. The second trough 372 may also be defined relative to the second peak 362 of the adjacent outlet chamber 320 (located on the opposite side of the second filter unit 130). In use, the second trough 372 is at the lowermost part of the intermediate chamber 330 (i.e., closer to the outlet end 106), and the second peak 362 is part of the adjacent outlet chamber 320 and is disposed at a higher position than the second trough 372 (i.e., closer to the inlet end 104).

[0083] The second filter unit 130 may define a second trough 372 by being angled and connected to at least one of the container walls 200, and may define a second peak 362 by being angled and connected to another container wall 200. For example, the second trough 372 may be defined by the second filter unit 130 and the rear wall 204. The angle (measured within the intermediate chamber 330) that defines the second trough 372 may be an acute angle. The second trough 372 may sometimes be referred to as the intermediate chamber trough. The second peak 362 may be defined by the second filter unit 130 and the front wall 202. The angle (measured within the intermediate chamber 330) that defines the second peak 362 may be an acute angle. When such an embodiment is used, the second filter unit 130 is in an inclined state inside the container 102.

[0084] The second filter unit 130 may be configured to filter a second substance from the fluid flowing from the intermediate chamber 330 to the outlet chamber 320. The second substance may be particles of a different size or type than the first substance. The second filter unit 130 may cooperate with the first filter unit 120 to filter substances from the fluid passing through the container 102 in stages. The staged filtration of substances can improve the yield of the desired fluid product and reduce the possibility of clogging by attempting to remove a large number of particles at once by the filter units 120, 130.

[0085] In some embodiments, the second substance may be particles of the same size or type as the first substance, and the second filter unit 130 is configured to capture or confine particles that are not intended to pass through the first filter unit 120. Similar to the first filter unit 120 and the first trough 342, the captured particles accumulated in the second filter unit 130 can be directed to the second trough 372. The second trough 372 is configured to receive the second substance. For example, the fluid in the intermediate chamber 330 can wash the second substance along the surface of the second filter unit 130 and direct it to the second trough 372. By inducing the second substance to accumulate in the second trough 372, more portions of the filter unit surface can be kept clear or unclogged, thus improving the filtration efficiency and / or reducing the time until the filter unit 130 (or the filtration device 100) needs to be cleaned or replaced.

[0086] The fluid that has passed through the second filter unit 130 enters the outlet chamber 320. The outlet chamber 320 is in fluid communication with the outlet port 110, whereby the fluid in the outlet chamber 320 is discharged from the filter container 102. The fluid in the outlet chamber 320 contains the product, i.e., the desired fluid product that has been substantially separated through the removal of particles by the filter units 120, 130. In some embodiments, even if a small amount of particles can pass through the filter units 120, 130, using the filtration device 100 significantly reduces the amount of particles in the filtered fluid compared to the unfiltered fluid. Further filtration may be performed to reduce the amount of remaining particles.

[0087] The outlet chamber 320 may extend from the outlet end 106 to the second filter unit 130 coupled to the container 102 and may be coupled, for example, to at least the front wall 202 and the rear wall 204. The second filter unit 130 may be coupled to the container 102 (such as the front wall 202) along the second front seam 360 and may be coupled to the container 102 (such as the rear wall 204) at the second back seam 370. The second front seam 360 and the second back seam 370 may extend to contact each other at the side ends 206, 208 in embodiments where the container 102 has a configuration similar to a saline bag. In some embodiments, the outlet chamber 320 may be substantially wedge-shaped. For example, the outlet chamber 320 may be shaped to gradually narrow from the outlet end 106 or near it towards the second peak 362.

[0088] The shape of the container 102 may be modified to space the second filter unit 130 from the front wall 202 / outlet port 110 as realized by the spacer system 220. The spacer system 220 can generally be classified as involving a structure that mechanically separates the second filter unit 130 and the front wall 202. In contrast, the shape of the container 102 is independent of the structure. However, the spacer system 220 can be used in combination with the modified shape of the container 102.

[0089] Figures 3A and 3B show embodiments of the container 102 with a modified shape.

[0090] The position of the second dorsal seam 370 may be adjusted to further separate the second filter unit 130 from the outlet port 110. Two exemplary positions of the second dorsal seam 370 are shown in FIG. 3A. The first position 370-1 is closer to the outlet end 106 compared to the second position 370-2. Thus, in the first position 370-1, the second filter unit 130 is closer to the outlet port 110 compared to the second position 370-2. The position indicated by the second filter unit 130 when the second dorsal seam 370 is in the second position 370-2 is shown in dashed lines. The second position 370-2 may be selected such that, in use, the outlet port 110 and the second filter unit 130 are arranged at an angle further apart from each other compared to the first position 370-1.

[0091] FIG. 3B shows an embodiment of the container 102 configured such that the front wall 202 bulges in a direction away from the second filter unit 130. FIG. 3B shows the front wall 202 in the bulged state 202-2. For comparison, the front wall 202 in the "normal" state 202-1 (as shown in FIG. 3A) is shown in dashed lines. In the bulged state 202-2, the front wall 202 may bulge between the second front seam 360 and the second dorsal seam 370. The front wall 202 may bulge between the second front seam 360 and the bottom seam 212. The amount of material between the second front seam 360 and the second dorsal seam 370 may be more, for example, than in the embodiment shown in FIG. 3A (the "normal" state 202-1). Thereby, the front wall 202 of the container 102 can bulge as in the bulged state 202-2.

[0092] The outlet port 110 is coupled to the front wall 202. The outlet port 110 has a mass sufficient to deform the front wall 202. For example, if the filtration device 100 is arranged such that fluid passes through the second filter unit 130 by gravity, the outlet port 110 is pulled downward by gravity, thereby pulling the front wall 202 downward and assuming a bulged state 202-2. The outlet port 110 may be disposed near the lower end 106 so as to pull the front wall 202 in a desired direction by its weight. When the fluid passes through the second filter unit 130 and enters the outlet chamber 320, due to the weight of the fluid, the front wall 202 may be pulled downward and assume a bulged state 202-2.

[0093] In the bulged state 202-2, the front wall 202 (and / or the outlet port 110) is separated from the second filter unit 130. Even if the second filter unit 130 is deflected by the fluid accumulated in the second filter unit 130, the front wall 202 (and / or the outlet port 110) still remains separated from the second filter unit 130, thereby keeping the holes of the second filter unit 130 unclogged or substantially clear, and the fluid can continue to pass through the second filter unit 130 and enter the outlet chamber 320.

[0094] By providing additional outlet ports 110, a further discharge location for the fluid in the outlet chamber 320 can be provided. In some embodiments, the outlet ports 110 may be arranged generally perpendicular to the deflection direction of the filter unit 130, thereby enabling the filter unit 130 to substantially not obstruct the outlet ports 110 even in a deflected state.

[0095] Since the first filter unit 120 and the second filter unit 130 are connected to the container 102 in a spaced-apart arrangement, there is a front seam distance between the first front seam 340 and the second front seam 360, and a back seam distance between the first back seam 350 and the second back seam 370. The distance of the front seam and the distance of the back seam may or may not be equal. In some embodiments, the front seam distance and / or the back seam distance is in the range of 0.5 inches to 5 inches. In some embodiments, the front seam distance and / or the back seam distance is about 1 inch.

[0096] In some embodiments, the filter units 120 and 130 are substantially planar (flat), and thus, when the front seam distance and the back seam distance are equal, the filter units 120, 130 are parallel. Thereby, the intermediate chamber 330 may have a square or rectangular cross-section.

[0097] In some embodiments, the filter units 120, 130 are substantially planar (flat), and thus, when the front seam distance and the back seam distance are not equal, the filter units 120, 130 are not parallel to each other (as shown in FIGS. 2A - 2D and FIGS. 3A, 3B). When the filter units 120, 130 are not parallel to each other, the filter units 120, 130 contact over most of their surface areas, reducing the possibility of blocking each other. For example, if the filter is a flexible bag bent into a curved shape, the filter units 120, 130 may contact only at a specific point rather than over most of their surface areas (even if there is a gap or they are in a non-parallel arrangement).

[0098] As shown in FIGS. 2A-2D and FIGS. 3A and 3B, the filter units 120, 130 may be angled or tilted relative to each other so as to generally form a "V" shape. The filter units 120, 130 may be angled or tilted relative to each other such that the cross-section of the intermediate chamber 330 is generally triangular, trapezoidal, or trapezoid-like in shape. In some embodiments, the minimum front seam distance is shorter than the minimum back seam distance.

[0099] The inclination of the filter units 120, 130 means that the first filter unit 120 and the second filter unit 130 are angled relative to each other, i.e., not parallel (e.g., as shown in FIGS. 2A-2D and FIGS. 3A and 3B). By combining with the spacing between the filter units 120 and 130, this angle reduces the possibility that the filter units 120 and 130 contact (and stick) to each other over most of their filter surfaces, particularly in embodiments where the container 102 is flexible. Thereby, clogging and / or damage of the filter mesh can be reduced. The inventors have found that when the filter meshes are arranged back-to-back, a depth is created that causes clogging, retains some cells, and as a result, the concentration may change and the yield may be affected. The "depth" in this context refers to the effective thickness of the filter that occurs when the first filter unit 120 and the second filter unit 130 contact or are very close to each other and the holes of one filter unit are blocked by the hole-defining structure of the other filter unit. For example, if the first filter unit 120 has a larger pore size than the second filter unit 130, arranging the filter units 120 and 130 close to each other may result in the first filter unit 120 having substantially the same pore size as the second filter unit 130. Therefore, the spacing and angle between the filter units 120, 130 can improve the flow through each filter and increase the yield.

[0100] In some embodiments, the filter units 120, 130 are conical (or triangular in cross-section) with equal front seam distance and back seam distance. The conical or triangular filter may have a peak portion rising towards the center of the inlet chamber 310 and descending towards the container wall 200 to define a first trough 342. Alternatively, the conical filter may define a first trough 342 near the center of the inlet chamber 310 and rise towards the container wall 200 to define a first peak 352. A similar configuration may be applied when defining a second peak 362 and a second trough 372 with respect to the intermediate chamber 330 and the outlet chamber 320.

[0101] To reduce the possibility that the filter units 120, 130 contact and block each other, the filter units 120, 130 may not be fitted together. For example, the first filter unit 120 does not extend inside or in the direction of the second filter unit 130, and vice versa.

[0102] System and Kit FIG. 4 shows a system 400 for filtering cells. The system may be supplied as a kit, in which case the system 400 is at least partially disassembled. The kit includes the filtration device 100 shown in FIGS. 1 - 3, an inlet conduit 500 (FIG. 5), and an outlet conduit 600 (FIG. 6).

[0103] The inlet conduit 500 includes a filling tube 510 adapted to couple to the inlet port 108 of the filtration device 100 to enable fluid communication with the inlet chamber 310. In use, the filling tube 510 can receive fluid from a fluid source 520 and direct it to the inlet chamber 310. The filling tube 510 may be connected to the inlet port 108 via an inlet coupling 530. The filling tube 510 may be in fluid communication with an inlet valve, which can be used to permit, block, and / or control the flow of fluid into the inlet chamber 310. In some embodiments, the inlet coupling 530 includes an inlet valve.

[0104] The outlet conduit 600 comprises a drain tube 610 adapted to couple to the outlet port 110 of the filtration device 100, enabling fluid communication with the outlet chamber 320. In use, the drain tube 610 can receive a product (filtered fluid) which can be a fluid from which the first and second substances have been removed, and direct it to another location such as one or more vials 620. The drain tube 610 can be connected to the outlet port 110 via an outlet coupling 630. The drain tube 610 may be in fluid communication with an outlet valve which can be used to permit, block, and / or control the flow of the filtered fluid from the outlet chamber 320. In some embodiments, the outlet coupling 630 includes the outlet valve.

[0105] In some applications of the system 400, it may include receiving fluid from multiple sources. In such situations, the fluids may be the same fluid. For example, to ensure a continuous supply of fluid passing through the filtration device 100, two fluid sources 520 may be connected to the system 400, and when the first fluid source 520A (e.g., a tank) becomes empty, the second fluid source 520B can supply fluid, minimizing / eliminating interruptions in supply. In another situation, the system 400 can receive two (or more) different types of fluids. This can enable mixing of the two fluids during or immediately before filtration. For example, when a cell composition is being prepared for filtration, the first fluid source 520A may contain cells and a formulation buffer (e.g., 2X formulation buffer), and the second fluid source 520B may contain a cryoprotectant to protect the cells from damage during cryopreservation. As an example, the cryoprotectant can be DMSO, but DMSO exhibits some cytotoxicity. Therefore, the cryoprotectant can be added immediately before filtration to minimize the time the cells are exposed to DMSO and ensure time for filling the filtered cell suspension into vials for freezing.

[0106] Accordingly, in some embodiments, the inlet conduit 500 may further include a first inlet tube 540 and a second inlet tube 550. The first inlet tube 540 is adapted to receive a first fluid from a first fluid source 520A, and the second inlet tube 550 is adapted to receive a second fluid from a second fluid source 520B, and the first fluid and the second fluid are different from each other. In some embodiments, the first inlet tube 540 may be in fluid communication with a first supply valve, and the second inlet tube 550 may be in fluid communication with a second supply valve. The first supply valve and the second supply valve can be used to permit, block, and / or control the flow of fluid from the fluid sources 520A, 520B.

[0107] The inlet tubes 540, 550 are configured to be in fluid communication with the filling tube 510. In some embodiments, the inlet conduit 500 includes a manifold 560. The filling tube 510 is configured to be in fluid communication with the manifold 560 and is adapted to receive fluid from at least one of the first and second inlet tubes 540, 550. The manifold 560 may be a Y-shaped or T-shaped connector through which the first fluid and the second fluid can flow from the inlet tubes 540, 550 into the filling tube 510. Accordingly, the inlet tubes 540, 550 and the filling tube 510 may form a Y-shape or a T-shape.

[0108] The various tubes 510, 540, 550, 610 may have an inner diameter in the range of 1 / 8 inch to 3 / 4 inch. The various tubes 510, 540, 550, 610 may have an outer diameter in the range of 1 / 4 inch to 1 inch. For example, in some embodiments, at least one of the tubes 510, 540, 550, 610 has an inner diameter of 1 / 8 inch and an outer diameter of 1 / 4 inch. However, other sizes of tubes may be used depending on the fluid flow characteristics (e.g., viscosity) and / or the desired throughput (flow rate, yield quality) of the system 400.

[0109] One or more of the components of the kit (filtration device 100, inlet conduit 500, and outlet conduit 600) can be made of DMSO-compatible plastic. In some embodiments, all components of the kit can be made of DMSO-compatible plastic. For example, DMSO-compatible plastic can be a plastic containing one or more of polyethylene terephthalate (PET), high density polyethylene (HDPE), low density polyethylene (LDPE), polytetrafluoroethylene (PTFE), polypropylene (PP), polyvinyl chloride (PVC), thermoplastic elastomer (TPE).

[0110] One or more of the components of the kit can include sterile weld components, such as for connecting filter units 120, 130 to container 102. System 400, the kit, and / or filtration device 100 can comply with USP 788.

[0111] The kit can further include a pump that generates pressure within system 400 to draw fluid through filtration device 100. The pump can be connected to outlet conduit 600 and can be used to assist in priming (removing air bubbles) of system 400. When fluid begins to flow into container 102, the pump operates to suck out the air within container 102, creating a pressure difference to facilitate the fluid passing through filter units 120, 130, and chamber 300. Removal of air can be preferred over introduction of air (through inlet conduit 500). This is because when air is introduced into system 400, air bubbles can form in the filtered fluid, and the amount / volume of the fluid obtained by filtration can decrease.

[0112] Operation In some embodiments, the fluid can flow through filtration device 100 by gravity without mechanical assistance. In use, container 102 is placed in an upright position such that inlet port 108 is higher than outlet port 110. In some embodiments, container 102 includes a flange 112 at inlet end 104. Flange 112 can define an opening 114 or connector for suspending container 102 from a hook to maintain the upright position.

[0113] In the upright position, the first filter unit 120 forms the inclined bottom of the inlet chamber 310 (the inclined lid of the intermediate chamber 330), and the second filter unit 130 forms the inclined bottom of the intermediate chamber 330 (the inclined lid of the outlet chamber 320). In the upright position, the flow of fluid passing through the filter units 120, 130 is assisted by gravity.

[0114] Before use, the system 400 needs to be "primed". Priming refers to removing air bubbles within the system 400 (such as in the inlet conduit 500 and the outlet conduit 600). To remove air bubbles, the inlet conduit 500 and the outlet conduit 600 can be manually and mechanically operated. Optionally, the container 102 can be compressed to expel the air within the container 102 and move the air bubbles along the inlet conduit 500 and the outlet conduit 600. During operation, the filtration device 100 may require intermittent priming.

[0115] In some embodiments, the filtration device 100 can be connected to a pump, and the pump generates pressure within the system 400 to draw fluid into the filtration device 100. The pump can be particularly useful when the filtration device 100 is used in a configuration where gravity is insufficient or has an adverse effect on the flow of fluid from the inlet port 108 to the outlet port 110. For example, when the heights of the inlet port 108 and the outlet port 110 are the same or nearly the same, the pump can assist the flow of fluid. The pump can be connected to the outlet conduit 600. The pump can assist in priming.

[0116] When fluid flows into the inlet chamber 310, the fluid contacts the inclined bottom of the inlet chamber 310 (the first filter unit 120). The first filter unit 120 prevents the first substance from passing through, and the first substance can be washed away into the first trough 342. The fluid passing through the first filter unit 120 may be referred to as the first filtered fluid.

[0117] The first filtered fluid passes through the first filter unit 120 and enters the intermediate chamber 330. The first filtered fluid contacts the inclined base (the second filter unit 130) of the intermediate chamber 330 and is filtered through the second filter unit 130. The second filter unit 130 is inclined at an angle different from that of the first filter unit 120 in some embodiments. The second substance is prevented from passing through the second filter unit 130 and the second substance can be washed into the second trough 372. The fluid passing through the second filter unit 130 may be referred to as the second filtered fluid.

[0118] The second filtered fluid passes through the second filter unit 130 and enters the outlet chamber 320. The second filtered fluid collects in the outlet chamber 320 and is discharged through the outlet port 110.

[0119] The filtration device 100 and kits containing the same can be used to filter various fluids such as stem cell culture media, cell compositions, and harvested cells suspended in cell culture media or another suitable buffer.

[0120] Some embodiments of the present disclosure relate to a method of filtering a stem cell culture medium. The method includes using the filtration device 100. The method may further include using a kit, and the stem cell culture medium is supplied through the inlet conduit 500 and discharged through the outlet conduit 600 after passing through the filtration device 100. The outlet conduit 600 can be used to transport the filtered stem cell culture medium for dispensing into at least one vial 620.

[0121] Some embodiments of the present disclosure relate to a method of filtering harvested cells in suspension. The harvested cells may coexist with an enzyme for detachment or a formulation reagent for filling. The method includes using the filtration device 100. The method may further include using a kit. The outlet conduit 600 can be used to transport the filtered harvested cells for dispensing into at least one vial 620.

[0122] Some embodiments of the present disclosure relate to a method for purifying a cell composition. The method includes reducing visible particles and / or cell aggregates by passing cultured cells through a double screen mesh filter such as filtration device 100. A kit can be used to carry out this method. The outlet conduit 600 can transport the purified cell composition for dispensing into at least one vial 620.

[0123] The first filter unit 120 of the filtration device 100 may have an average pore size in the range of 130 μm to 170 μm, for example, 150 μm. The second filter unit 130 of the filtration device 100 may have an average pore size in the range of 20 μm to 60 μm, for example, 40 μm. The purified cell composition may be substantially free of visible particles. The inspection process can be carried out visually by a trained and qualified operator based on, for example, USP 790 and USP 1790. A light box with a black and white background can be used for carrying out the inspection. In this inspection, it may not be necessary to use magnified or polarized illumination.

[0124] The cultured cells can be provided in a serum-free cell culture medium. In some embodiments, the cultured cells are provided in a cell culture medium containing serum such as fetal bovine serum (FBS). As is well known to those skilled in cell culture, cell culture media contain various components to support cell growth, which mainly depend on the cells being cultured. Exemplary cell culture media and their components are described below.

[0125] In some embodiments of this method, the cells are cultured and grown.

[0126] In some embodiments of this method, the cells are mesenchymal progenitor cells or stem cells (MLPSCs).

[0127] After passing the cells through the filtration device 100, the recovery rate of viable cell concentration can be 60% to 100%. For example, a 100% recovery rate of viable cells means that the product / fluid after filtration contains all the cells that were contained in the fluid introduced into the inlet chamber 310. A 60% recovery rate indicates that 40% of the cells contained in the fluid introduced into the inlet chamber 310 remain within the filtration device 100. Those cells are captured, for example, in the first filter unit 120 together with the first substance or in the second filter unit 130 together with the second substance. In some embodiments, the recovery rate is 70% to 90%.

[0128] In some embodiments of this method, the D90 of the purified cell composition is less than 150 μm, preferably less than 100 μm, more preferably less than 50 μm.

[0129] Relationship between chamber volume and filter unit surface area When determining the sizes of the chamber 300 and the filter units 120, 130, it is necessary to consider the relationship between the chamber volume and the filter unit surface area. If the surface area of the filter unit is too small relative to the volume of the chamber supplying the filter unit, the filter unit is likely to function as a bottleneck or choke point. This can reduce the speed of the entire filtration process and affect the yield. The bottleneck can cause clogging of the filter unit.

[0130] In some embodiments, the filter unit is disposed inclined inside the filter container 102. By inclining the filter unit with respect to the wall 200 of the container 102, a filter unit with a larger surface area can be installed inside the container 102 compared to a configuration where the filter unit is not inclined.

[0131] If the filter unit is not tilted inside the container 102, it may be necessary to increase the size of the container 102 to accommodate the filter unit. Enlarging the size of the container 102 may affect the chamber volume, but the size of the container 102 should be in an appropriate ratio to the filter surface area in order to reduce the possibility that the filter unit becomes a bottleneck or choke point, as described above. When using the container 102 in a small space or a place with a complex shape, it may be difficult to enlarge the size of the container 102.

[0132] To promote the flow of fluid, it is also necessary to consider the relative volumes of the inlet chamber 310, the intermediate chamber 330, and the outlet chamber 320. If the volume of the next chamber (e.g., the intermediate chamber 330) is larger than the volume of the previous chamber (e.g., the inlet chamber 310 that supplies the intermediate chamber 330), it usually does not have an adverse effect on the fluid flow. However, if the volume of the next chamber is too small compared to the volume of the previous chamber, the speed at which the fluid flows into the next chamber may exceed the outflow speed. This may result in a bottleneck or choke point. Also, the pore size of the filter unit and the fluid flow rate through the inlet port 108 and the outlet port 110 also affect the movement speed of the fluid passing through the various chambers 300 in the filter, and as a result, affect the size of the chamber volume.

[0133] In some embodiments, the volumes of the inlet chamber 310, the intermediate chamber 330, and the outlet chamber 320 are all approximately equal. Therefore, the chamber volume can be represented by a ratio of 1:1:1.

[0134] In some embodiments, the volumes of two of the inlet chamber 310, the intermediate chamber 330, and the outlet chamber 320 are approximately equal. The volume of the other chamber may be smaller than at least one of the volumes of the other chambers. For example, the volume of the other chamber may be 30% - 70% of the volume of one of the other chambers.

[0135] In some embodiments, the intermediate chamber 330 is twice the size of the inlet chamber 310 and the outlet chamber 320. Thus, the volume of the chambers can be represented by a ratio of 1:2:1 (inlet:intermediate:outlet).

[0136] In some embodiments, the volumes of the intermediate chamber 330 and the outlet chamber 320 are approximately equal. The volume of the inlet chamber 310 may be smaller than theirs. The volume of the chambers can be represented by a ratio of 1:2:2 (inlet:intermediate:outlet).

[0137] In some embodiments, the volumes of the inlet chamber 310 and the intermediate chamber 330 are approximately equal. The volume of the outlet chamber 320 may be smaller than theirs. The volume of the chambers can be represented by a ratio of 2:2:1 (inlet:intermediate:outlet).

[0138] In some embodiments, the inlet chamber 310, the intermediate chamber 330, and the outlet chamber 320 have different volumes from each other. For example, the volume of the intermediate chamber 330 can be three times the size of the inlet chamber 310, and the volume of the outlet chamber 320 can be twice the size of the inlet chamber 310. Thus, the volume of the chambers can be represented by a ratio of 1:3:2.

[0139] In some embodiments, the intermediate chamber 330 has the largest volume among all the chambers. By making the intermediate chamber 330 the chamber with the largest volume, the fluid that has passed through the first filter unit 120 can accumulate in the intermediate chamber 330 before passing through the second filter unit 130, thereby smoothing the flow of the fluid passing through the filtration device 100. In some embodiments, the total volume of the container 102 is 1.5 liters, which can allow approximately 2 liters of fluid to pass while minimizing fouling and clogging of the filter units 120, 130. In some embodiments, the total volume of the container 102 is 2.0 liters.

[0140] In an embodiment where the outlet chamber 320 is smaller than the intermediate chamber 330, the flow rate of the fluid discharged through the outlet port 110 can be controlled so that excessive fluid does not accumulate in the outlet chamber 320. Thereby, the risk that excessive fluid in the outlet chamber 320 affects the fluid flow from the intermediate chamber 330 is reduced.

[0141] Method In one embodiment, the present disclosure relates to a method of filtering a cell culture medium containing cells, the method including passing the cells through a filter described herein. For example, the cells can be passed through the screen mesh filter described above. When using the method of the present disclosure, there is no particular limitation on the cell type, and in some examples, it is related to both differentiated and undifferentiated cells. As an example, the cells are mesenchymal or progenitor cells (MLPSCs). In this example, the method of the present disclosure can be used to filter a stem cell culture medium. For example, MLPSCs are cultured according to the method described below and then filtered by passing the cells through a filter described herein. In one example, the cells are provided in a stem cell culture medium. In one example, the stem cell culture medium is purified or partially purified before the resulting composition containing the cells is passed through a filter described herein. For example, the stem cell culture medium containing cells can be centrifuged to remove the culture medium from the cells. The cells can then be resuspended in an appropriate buffer or resuspension medium. In this example, centrifugation and resuspension may be repeated multiple times to wash the cells. The centrifuged cells are then resuspended in an appropriate buffer or resuspension medium and passed through a filter disclosed herein.

[0142] In certain embodiments, cells passed through the filters disclosed herein may be referred to as a purified cell composition. In one example, the purified cell composition is characterized by specific structural features that can be specifically identified via visual inspection or other analytical methods. For example, the purified cell composition may substantially lack visible particles. In another example, the purified cell composition is characterized by the size of the aggregates. As used herein, "aggregate" means the totality of a plurality of individual cells within a cluster grouped by one or more adhesion properties including aggregation, agglutination, and adhesion. As used herein, "agglutination" means the tendency of cells to aggregate. In one example, 90% of the diameter (D 90 ) of a cell population / aggregate (e.g., a stem cell population / aggregate) is less than 150 μm. For example, the purified stem cell composition included in the present disclosure may have a D 90 of less than 150 μm. In another example, D 90 is less than 100 μm. In another example, D 90 is less than 50 μm. For example, D 90 can be between 50 μm and 150 μm. In another example, D 90 can be between 50 μm and 100 μm.

[0143] In certain embodiments, a viable cell concentration of 60% to 100% is provided by passing a cell or cell culture medium, or a resuspension medium containing them, through the filter described herein. In some embodiments, the viable cell recovery rate is 70% to 90%. In one example, the viable cell recovery rate is determined relative to the number of viable cells in the composition before passing through the filter described herein. In another example, the percentage of the recovery rate is determined relative to the total number of cells in the purified cell composition (i.e., after filtration). Various standard methods for determining cell viability are known in the art. In one example, automated cell counting is used. Such methods may use techniques and devices based on Coulter counting or flow cytometry. These systems generally rely on counting the number of particles of a specific size by electrical or optical detection, and in some cases, a dye (or the like) can also be incorporated to distinguish dead cells from live cells. In another example, semi - automated or manual cell counting can be used to determine cell viability. Such methods generally involve using an assay containing well - known substances such as trypan blue or propidium iodide (PI) to label dead cells and then counting the number of dead cells relative to the number of live cells in the sample. As will be understood by those skilled in the art, cell viability is usually evaluated in a sample representative of a larger composition, and the results from the representative sample are extrapolated to provide an overall level of cell viability for the larger composition.

[0144] Mesenchymal progenitor cells or stem cells As used herein, the term "mesenchymal progenitor cells or stem cells (MLPSC)" refers to undifferentiated pluripotent cells that have the ability to self - replicate while maintaining pluripotency and the ability to differentiate into a number of cell types, either of mesenchymal origin, such as osteoblasts, chondrocytes, adipocytes, stromal cells, fibroblasts and tendons, or of non - mesodermal origin, such as hepatocytes, neurons and epithelial cells. To avoid misunderstanding, "mesenchymal progenitor cells" refers to cells that can differentiate into mesenchymal cells such as bone, cartilage, muscle and fat cells, as well as cells that can differentiate into fibrous connective tissue.

[0145] The term "mesenchymal progenitor cells or stem cells" includes both parental cells and their undifferentiated progeny. This term also includes mesenchymal progenitor cells, multipotent stromal cells, mesenchymal stem cells (MSCs), perivascular mesenchymal progenitor cells, and their undifferentiated progeny.

[0146] Mesenchymal progenitor cells or stem cells can be autologous, allogeneic, xenogeneic, syngeneic, or isogeneic. Autologous cells are isolated from the same individual into which they are re-transplanted. Allogeneic cells are isolated from a donor of the same species. Xenogeneic cells are isolated from a donor of a different species. Syngeneic or isogeneic cells are isolated from genetically identical organisms, such as twins, clones, or highly inbred laboratory animal models.

[0147] In one example, mesenchymal progenitor cells or stem cells are allogeneic. In one example, allogeneic mesenchymal progenitor cells or stem cells are cultured and cryopreserved.

[0148] Mesenchymal progenitor cells or stem cells are mainly present in the bone marrow, but have also been shown to be present in a variety of host tissues, including, for example, umbilical cord blood and cord, adult peripheral blood, adipose tissue, trabecular bone, and dental pulp. They are also found in the skin, spleen, pancreas, brain, kidney, liver, heart, retina, brain, hair follicles, intestine, lung, lymph nodes, thymus, ligaments, tendons, skeletal muscle, dermis, and periosteum, and can differentiate into germ cell lineages such as mesoderm and / or endoderm and / or ectoderm. Thus, mesenchymal progenitor cells or stem cells can differentiate into a number of cell types, including but not limited to adipose, bone, cartilage, elastic, muscle, and fibrous connective tissues. The specific lineage-commitment and differentiation pathways into which these cells enter depend on mechanical influences and / or endogenous bioactive factors, such as growth factors, cytokines, and / or various influences from the local microenvironmental conditions established by the host tissue.

[0149] As used herein, the terms "enriched", "enrichment" or variations thereof are used to describe a population of cells in which the proportion of one particular cell type or the proportion of the number of several particular cell types is increased compared to a population of untreated cells (e.g., cells in their native environment). In one example, a population enriched in mesenchymal progenitor cells or stem cells contains at least about 0.1%, or 0.5%, or 1%, or 2%, or 5%, or 10%, or 15%, or 20%, or 25%, or 30%, or 50%, or 75% mesenchymal progenitor cells or stem cells. In this regard, the term "population of cells enriched in mesenchymal progenitor cells or stem cells" is construed to provide an express underpinning of the term "population of cells containing X% mesenchymal progenitor cells or stem cells", where X% is the percentage described herein. Mesenchymal progenitor cells or stem cells can, in some instances, form clonogenic colonies; for example, CFU-F (fibroblasts) or a subset thereof (e.g., 50% or 60% or 70% or 70% or 90% or 95%) can have this activity.

[0150] In one example of the present disclosure, the mesenchymal progenitor cells or stem cells are mesenchymal stem cells (MSCs). The MSCs can be a homogeneous composition or a mixed cell population enriched in MSCs. A homogeneous MSC composition can be obtained by culturing adherent bone marrow cells or periosteal cells, and the MSCs can be identified by specific cell surface markers that are identified by a proprietary monoclonal antibody. Methods for obtaining a cell population enriched in MSCs are described, for example, in U.S. Patent No. 5,486,359. Alternative sources of MSCs include, but are not limited to, blood, skin, cord blood, muscle, fat, bone, perichondrium. In one example, the MSCs are allogeneic. In one example, the MSCs are cryopreserved. In one example, the MSCs are cultured and expanded and cryopreserved.

[0151] In another example, the mesenchymal progenitor cells or stem cells are CD29+, CD54+, CD73+, CD90+, CD102+, CD105+, CD106+, CD166+, MHC1+ MSCs.

[0152] In one example, mesenchymal progenitor cells or stem cells are cultured and expanded from a population of MSCs that express markers including CD73, CD90, CD105, CD166 and do not express hematopoietic cell surface antigens such as CD45 and CD31. For example, mesenchymal progenitor cells or stem cells can be cultured and expanded from a population of MSCs that are CD73+, CD90+, CD105+, CD166+, CD45−, CD31−. In one example, the population of MSCs is further characterized by a low level of major histocompatibility complex (MHC) class I. In another example, MSCs are negative for major histocompatibility complex class II molecules and negative for co-stimulatory molecules CD40, CD80, CD86. In one example, the culture expansion process includes 5 passages.

[0153] Isolated or enriched mesenchymal progenitor cells or stem cells can be expanded in vitro by culture. Isolated or enriched mesenchymal progenitor cells or stem cells can be cryopreserved, thawed, and then expanded in vitro by culture.

[0154] In one example, isolated or enriched mesenchymal progenitor cells or stem cells are seeded at about 50,000 viable cells per cm in a culture medium (serum-free or serum-supplemented), such as alpha minimum essential medium (αMEM) supplemented with 5% fetal bovine serum (FBS) and glutamine. 2 and allowed to attach to the culture vessel overnight at 37°C, 20% O 2 . Then, the culture medium is changed and / or adjusted as needed, and the cells are cultured for an additional 68 - 72 hours at 37°C, 5% O 2 .

[0155] As will be understood by those skilled in the art, cultured mesenchymal progenitor cells or stem cells have a different phenotype from in vivo cells. For example, in one embodiment, they express one or more of the markers CD44, NG2, DC146, and CD140b. Cultured mesenchymal progenitor cells or stem cells are also biologically different from in vivo cells and have a higher proliferation rate than most of the in vivo quiescent (resting) cells.

[0156] In one example, the population of cells is enriched from a cell preparation comprising STRO-1+ cells in a selectable form. In this regard, the term "selectable form" is understood to mean that the cells express a marker (e.g., a cell surface marker) that enables the selection of STRO-1+ cells. The marker can be STRO-1, but it does not have to be. For example, as described and / or exemplified herein, cells that express STRO-2 and / or STRO-3 (TNAP) and / or STRO-4 and / or VCAM-1 and / or CD146 and / or 3G5 (e.g., mesenchymal progenitor cells) also express STRO-1 (which can be STRO-1bright). Thus, the designation that a cell is STRO-1+ does not mean that the cell is selected by STRO-1 expression alone. In one example, the cells are selected based at least on STRO-3 expression, e.g., they are STRO-3+ (TNAP+).

[0157] References to the selection of a cell or population thereof do not necessarily require selection from a particular tissue source. As described herein, STRO-1+ cells can be selected, isolated, or enriched from a wide variety of sources. That said, in some examples, these terms provide support for the selection from any tissue comprising STRO-1+ cells (e.g., mesenchymal progenitor cells), or vascular tissue, or tissue comprising pericyte cells (e.g., STRO-1+ pericytes), or any one or more of the tissues enumerated herein.

[0158] In one example, the cells used in the present disclosure express one or more markers individually or collectively selected from the group consisting of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β), CD45+, CD146+, 3G5+, or any combination thereof.

[0159] "Individually" means that the present disclosure separately encompasses the recited marker or markers, and that even if the individual marker or markers cannot be separately recited herein, the appended claims can define such marker or markers separately and divisibly from each other.

[0160] "Collectively" means that the present disclosure encompasses any number or combination of the recited marker or markers, and that even if such number or combination of marker or markers cannot be specifically recited herein, the appended claims can define such combination or sub-combination separately and divisibly from any other combination of marker or markers.

[0161] As used herein, the term "TNAP" is intended to encompass all isoforms of tissue-nonspecific alkaline phosphatase. For example, the term encompasses liver isoform (LAP), bone isoform (BAP), and kidney isoform (KAP). In one example, TNAP is BAP. In one example, TNAP as used herein refers to a molecule that can bind to the STRO-3 antibody produced by the hybridoma cell line deposited with the ATCC on December 19, 2005, under the terms of the Budapest Treaty, with the deposit accession number PTA-7282.

[0162] Furthermore, in one example, STRO-1+ cells can give rise to clonogenic CFU-F.

[0163] In one example, a significant proportion of STRO-1+ cells can differentiate into at least two different germ cell lines. Non-limiting examples of lineages to which STRO-1+ cells can commit include: bone progenitor cells; hepatocyte precursors that are pluripotent for bile duct epithelial cells and hepatocytes; neural-restricted cells that can give rise to glial cell precursors that develop into oligodendrocytes and astrocytes; neuron precursors that develop into neurons; precursors of cardiomyocytes and cardiomyocytes, glucose-responsive insulin-secreting pancreatic beta cell lines. Other lineages include, but are not limited to, odontoblasts, dentin-producing cells and chondrocytes, and the following: retinal pigment epithelial cells, fibroblasts, skin cells such as keratinocytes, dendritic cells, hair follicle cells, renal tubular epithelial cells, smooth and skeletal muscle cells, testicular precursors, vascular endothelial cells, tendons, ligaments, cartilage, adipocytes, fibroblasts, bone marrow stroma, cardiomyocytes, smooth muscle, skeletal muscle, pericytes, blood vessels, epithelium, glia, neurons, astrocytes and oligodendrocyte precursor cells.

[0164] In one example, mesenchymal progenitor cells or stem cells are obtained from a single donor or multiple donors, the donor samples or mesenchymal progenitor cells or stem cells are then pooled and then cultured and expanded.

[0165] The mesenchymal progenitor cells or stem cells encompassed by the present disclosure can also be cryopreserved before administration to a subject. In one example, the mesenchymal progenitor cells or stem cells are cultured and expanded and cryopreserved before administration to a subject.

[0166] Culturing and expansion of cells In one example, mesenchymal progenitor cells or stem cells are cultured and expanded. "Cultured and expanded" mesenchymal progenitor cells or stem cell cultures are distinguished from freshly isolated cells in that they are cultured and passaged in a cell culture medium (i.e., subcultured). In one example, the cultured and expanded mesenchymal progenitor cells or stem cells are expanded by about 4 to 10 passages. In one example, the mesenchymal progenitor cells or stem cells are subcultured at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 times. For example, the mesenchymal progenitor cells or stem cells can be subcultured and expanded at least 5 times. In one example, the mesenchymal progenitor cells or stem cells can be subcultured and expanded at least 5 to 10 times. In one example, the mesenchymal progenitor cells or stem cells can be subcultured and expanded at least 5 to 8 times. In one example, the mesenchymal progenitor cells or stem cells can be subcultured and expanded at least 5 to 7 times. In one example, the mesenchymal progenitor cells or stem cells can be cultured and expanded beyond 10 passages. In another example, the mesenchymal progenitor cells or stem cells can be cultured and expanded beyond 7 passages. In these examples, the stem cells can be cultured and expanded prior to cryopreservation to provide an intermediate cryopreserved MLPSC population. In one embodiment, the compositions of the present disclosure are prepared from the intermediate cryopreserved MLPSC population. For example, the intermediate cryopreserved MLPSC population can be further cultured and expanded prior to administration. Thus, in one example, the mesenchymal progenitor cells or stem cells are cultured and expanded and cryopreserved. In one embodiment of these examples, the mesenchymal progenitor cells or stem cells can be obtained from a single donor or multiple donors, the donor samples or mesenchymal progenitor cells or stem cells are then pooled and then cultured and expanded. In one example, the culture expansion process includes: i. increasing the number of viable cells by passage expansion, including establishing a primary culture of the isolated mesenchymal progenitor cells or stem cells and then successively establishing a first non-primary (P1) culture of the mesenchymal progenitor cells or stem cells isolated from the previous culture by passage expansion to provide a preparation of at least about 1 billion viable cells; ii. Propagating an isolated P1 culture of mesenchymal progenitor cells or stem cells by subculturing it into a second non-primary (P2) culture of mesenchymal progenitor cells or stem cells; and, iii. Preparing and cryopreserving an in-process intermediate mesenchymal progenitor cell or stem cell preparation obtained from the P2 culture of mesenchymal progenitor cells or stem cells; and, iv. Thawing the cryopreserved in-process intermediate mesenchymal progenitor cell or stem cell preparation and growing the in-process intermediate mesenchymal progenitor cell or stem cell preparation by subculture.

[0167] In one example, the expanded mesenchymal progenitor cell or stem cell preparation has an antigen profile and activity profile that includes: i. Less than about 0.75% CD45+ cells; ii. At least about 95% CD105+ cells; iii. At least about 95% CD166+ cells.

[0168] In one example, the expanded mesenchymal progenitor cell or stem cell preparation can inhibit IL2Rα expression by CD3 / CD28-activated PBMCs by at least about 30% compared to a control.

[0169] In one example, the cultured and expanded mesenchymal progenitor cells or stem cells are cultured and expanded for about 4 - 10 passages, the mesenchymal progenitor cells or stem cells are cryopreserved after at least 2 or 3 passages, and then further cultured and expanded. In one example, the mesenchymal progenitor cells or stem cells are cultured and expanded for at least 1, at least 2, at least 3, at least 4, at least 5 passages, cryopreserved, and then administered or cryopreserved again after further at least 1, at least 2, at least 3, at least 4, at least 5 passages of culture and expansion.

[0170] In one example, most of the mesenchymal progenitor cells or stem cells in the compositions of the present disclosure are of approximately the same generation number (i.e., the cells are within the range of about 1, about 2, about 3, or about 4 cell divisions of each other). In one example, the average number of cell divisions in the present composition is from about 20 to about 25. In one example, the average number of cell divisions in the present composition is from about 9 to about 13 (e.g., about 11 or about 11.2) divisions resulting from primary culture, plus about 1, about 2, about 3, or about 4 (e.g., about 2.5 per passage). Exemplary average numbers of cell divisions in the present composition are about 13.5, about 16, about 18.5, about 21, about 23.5, about 26, about 28.5, about 31, about 33.5, and about 36 when subjected to about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, and about 10 passages of proliferation, respectively.

[0171] The process of isolating and ex vivo expanding mesenchymal progenitor cells or stem cells can be carried out using any device and cell handling method known in the art. The various culture expansion embodiments of the present disclosure use steps that require manipulation of the cells, such as seeding, feeding, dissociation of adherent cultures, or washing steps. Any step of manipulating the cells has the potential to damage the cells. Mesenchymal progenitor cells or stem cells can generally withstand a certain degree of damage during preparation, but it is preferred to manipulate the cells with handling procedures and / or devices that appropriately perform the predetermined step(s) while minimizing damage to the cells.

[0172] In one example, the mesenchymal progenitor cells or stem cells are washed in a device comprising a cell source bag, a wash solution bag, a recirculation wash bag, a rotating membrane filter with inlet and outlet ports, a filtrate bag, a mixing zone, a final product bag for the washed cells, and appropriate tubing, as described, for example, in U.S. Patent No. 6,251,295, which is incorporated herein by reference.

[0173] In one example, the mesenchymal progenitor cell or stem cell composition according to the present disclosure is 95% homogeneous in that it is CD105 positive, CD166 positive, and CD45 negative. In one example, this homogeneity persists through ex vivo expansion, i.e., even through multiple population doublings. In one example, the composition contains at least one therapeutic dose of mesenchymal progenitor cells or stem cells, and the mesenchymal progenitor cells or stem cells contain less than about 1.25% CD45+ cells, at least about 95% CD105+ cells, and at least about 95% CD166+ cells. In one example, this homogeneity is maintained after cryopreservation and thawing, and the cells also generally have a viability of about 70% or more.

[0174] Cell culture medium The mesenchymal progenitor cells or stem cells disclosed herein can be cultured and expanded in various suitable growth media. As used in the context of the present disclosure, the term "medium" or "media" includes the components of the environment surrounding the cells. The medium contributes to and / or provides conditions suitable for growing the cells. The medium can be solid, liquid, gaseous, or a mixture of these phases and materials. The medium can include liquid growth media, as well as liquid media that do not support cell growth. The medium also includes gelatin media such as agar, agarose, gelatin, and collagen matrices. An exemplary gaseous medium includes the gas phase to which cells growing on a Petri dish or other solid or semi-solid support are exposed.

[0175] The cell culture medium used for culture and expansion contains all essential amino acids and may also contain non-essential amino acids. Generally, amino acids are classified into essential amino acids (Thr, Met, Val, Leu, Ile, Phe, Trp, Lys, His) and non-essential amino acids (Gly, Ala, Ser, Cys, Gln, Asn, Asp, Tyr, Arg, Pro).

[0176] One of ordinary skill in the art will understand that for optimal results, the basal medium must be appropriate for the cell line of interest. For example, it may be necessary to increase the level of glucose (or other energy source) in the basal medium or add glucose (or other energy source) during the culturing process if it is found that this energy source is depleted and thus limits growth. In one example, the dissolved oxygen (DO) level can also be controlled.

[0177] In one example, the cell culture medium contains human-derived additives. For example, human serum and human platelet cell lysate can be added to the cell culture medium. For example, the medium can contain 2 - 10% human serum. As an example, the medium contains 3% human serum by volume.

[0178] In one example, the cell culture medium contains only human-derived additives. Thus, in one example, the cell culture medium is xeno-free. To avoid misunderstanding, in these examples, the culture medium does not contain animal proteins.

[0179] In one example, the culture medium contains serum. In other examples, the culture medium is a culture medium that does not contain fetal bovine serum and contains growth factors that promote the proliferation of mesenchymal progenitor cells or stem cells. In one embodiment, the culture medium is a serum-free stem cell culture medium. In one example, the cell culture medium contains the following: Basal medium; Platelet-derived growth factor (PDGF); Fibroblast growth factor 2 (FGF2).

[0180] In one example, the culture medium contains platelet-derived growth factor (PDGF) and fibroblast growth factor 2 (FGF2), and the level of FGF2 is less than 5 ng / ml. For example, the FGF2 level can be 1 - 2 ng / ml.

[0181] In one example, the PDGF is PDGF-BB. In one example, the level of PDGF-BB is from about 1 ng / ml to 150 ng / ml. In another example, the level of PDGF-BB is from about 7.5 ng / ml to 20 ng / ml. In another example, the level of PDGF-BB is at least 10 ng / ml.

[0182] In other examples, additional factors can be added to the cell culture medium. In one example, the culture medium further contains EGF. EGF is a growth factor that stimulates cell proliferation by binding to its receptor EGFR. In one example, the level of EGF is from 0.1 to 7 ng / ml. For example, the level of EGF can be at least 5 ng / ml. In one example, the level of EGF is from about 2 to 5 ng / ml.

[0183] In the above examples, a reference amount of growth factor can be supplemented to a basal medium such as Alpha MEM or StemSpan™.

[0184] In other examples, additional factors can be added to the cell culture medium. For example, the cell culture medium can be supplemented with one or more stimulatory factors selected from the group consisting of epidermal growth factor (EGF), lα,25-dihydroxyvitamin D3 (1,25D), tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and stromal cell-derived factor 1α (SDF-1α).

[0185] In another example, the cell culture medium promotes the proliferation of stem cells while maintaining the stem cells in an undifferentiated state. Stem cells are considered undifferentiated when they are not committed to a particular differentiation lineage. As described above, stem cells exhibit morphological characteristics that distinguish them from differentiated cells. Furthermore, undifferentiated stem cells express genes that can be used as markers for detecting the differentiated state. The polypeptide product can also be used as a marker for detecting the differentiated state. Thus, one of ordinary skill in the art can readily determine whether the method of the present disclosure maintains stem cells in an undifferentiated state using conventional morphological analysis, genetic analysis, and / or proteomic analysis.

[0186] Prototype and experimental data The inventors tested the filtration device 100 with the configuration shown in FIGS. 1 to 3, in which the total volume of the container 102 was in the range of 1.5 liters to 2 liters. The first filter unit 120 had a pore diameter of 150 μm and a surface area of about 258 cm 2 . The pore diameter of the second filter unit 130 was 40 μm and the surface area was 180 cm 2 . The chamber volumes are summarized in Table 1 below.

[0187] [Table 1]

[0188] Considering the minimum and maximum chamber volumes in Table 1, the ratio of the chamber volume to the surface area of the filter unit is as follows. Inlet chamber: Minimum inlet chamber volume = 250 cm 3 Maximum inlet chamber volume = 800 cm 3 Therefore, the inlet chamber volume is in the range of about 0.9 (250 / 258) to about 3.2 (800 / 258) times the surface area of the first filter unit. Intermediate chamber: Minimum intermediate chamber volume = 500 cm 3 Maximum intermediate chamber volume = 1000 cm 3 Therefore, the intermediate chamber volume is in the range of about 1.9 (500 / 258) to about 4.0 (1000 / 258) times the surface area of the first filter unit. Minimum intermediate chamber volume = 500 cm 3 Maximum intermediate chamber volume = 1000 cm 3 Therefore, the intermediate chamber volume is in the range of about 2.7 (500 / 180) to about 5.6 (1000 / 180) times the surface area of the second filter unit. Outlet chamber: Minimum outlet chamber volume = 300 cm 3 Maximum outlet chamber volume = 800 cm 3 Therefore, the outlet chamber volume is in the range of about 1.6 (300 / 180) to about 4.5 (800 / 180) times the surface area of the second filter unit.

[0189] The filter units 120, 130 are connected to the container wall 200 in an inclined arrangement with respect to each other, forming a "V" shape when viewed in cross-section (as shown, for example, in FIGS. 2A - 2D and FIGS. 3A and 3B). The distance between the front seams 340, 360 (the portions where the first filter unit 120 and the second filter unit 130 are respectively coupled to the front wall 202 of the container 102) was about 1 inch.

[0190] To determine the filtration efficiency of the filtration device 100, the inventors used beads representing particles with a size greater than 40 μm and another type of beads representing cells. The beads representing particles with a size greater than 40 μm were Cosphoric red beads (catalog number UVPMS - BR - 1.20 45 - 53 μm (density 1.20 g / cc, size range 45 - 53 μm)). The beads used to represent cells were Cosphoric blue beads (catalog number BLPMS - 1.08 20 - 27 μm (density 1.08 g / cc, size range 20 - 27 μm)). Both types of beads were mixed in a matrix (MES Stop from Lonza Singapore) having the same density as the αMEM cell culture medium containing 10% DMSO (BloodStor 100).

[0191] The filtration device 100 was also tested using Cospheric red beads (45 - 53 μm) representing particles with a size greater than 40 μm alone in the BloodStor 100 matrix.

[0192] Some embodiments of the filtration device 100 were tested using cells in a BloodStor100 matrix representing a quenched cell suspension (1 L TrypLE + 4 L of v2.2 medium).

[0193] Some embodiments of the filtration device 100 were tested using cells concentrated by centrifugation at 400×g for 8 minutes at 2-8°C. The cells were resuspended in 400 mL of modified αMEM (SGTS-10533, Lonza) to represent the concentrated cell suspension.

[0194] Some embodiments of the filtration device 100 were tested using cells concentrated by centrifugation at 400×g for 8 minutes at 2-8°C. The cells were resuspended in 300 mL of cryopreservation medium containing Plasmalyte A, 5% human serum albumin, and 10% DMSO to represent a 1X formulation.

[0195] To determine the amount of cells filtered from the fluid (including beads in the matrix), two 1 mL samples were taken before and after filtration, respectively. The number of both types of beads in a 10 μL aliquot was counted using a glass hemocytometer or the trypan blue exclusion method. The filtration efficiency was determined by comparing the amount of beads measured before and after filtration. The weight of the fluid was measured before and after filtration to determine the amount of fluid retained in the filtration device 100.

[0196] The inventors confirmed that the filtration device 100 shown in FIGS. 1-3 successfully filtered beads representing particles larger than 40 μm without problem and passed cells (20-27 μm) without hindering their flow. Slightly red beads (representing particles larger than 40 μm) were observed in the post-filtration sample, but these were the same size as the 20-27 μm beads. Therefore, the number of 45-53 μm beads was considered "zero". The removal performance of particles and cells did not seem to be affected in the various matrices tested, especially those containing 10% DMSO.

[0197] To compare the performance, the inventors also tested a commercially available filtration device (Haemonetics SQ40S) having a 40 μm filter unit / screen. This filtration device successfully filtered beads representing particles larger than 40 μm in size without problems. However, the flow of beads representing cells (20 - 27 μm) was blocked, resulting in a loss of 21.9% of the small beads.

[0198] The inventors observed in some embodiments that a portion of the 20 - 27 μm beads (representing cells) remained attached inside the chamber 300. The matrix (including beads and cells) passed through the filter units 120, 130 with little resistance. The inventors observed that by allowing the filtration device 100 to accumulate approximately 1 / 3 or 1 / 2 of its volume capacity (about 300 - 500 mL), the amount of residual beads could be reduced. To enable the filtration device 100 to accumulate in this way, a clamp or valve was used to temporarily stop the flow of fluid from the filtration device 100 (e.g., a clamp or valve connected to the drain tube 610). When the filtration device 100 was filled from about 1 / 3 to 1 / 2, the outlet clamp / valve was removed. In some embodiments, it was observed that this improved the flow of fluid through the filtration device 100. The inventors observed in some embodiments that the accumulated fluid reduced the possibility that the wall 200 of the filtration device 100 would act as a bottleneck upon contact, thereby inducing the accumulation of residual beads. The accumulated fluid was observed in some embodiments to enable improved control of the purification process from a handling perspective.

[0199] It will be understood by those skilled in the art that numerous variations and / or modifications may be made to the above-described embodiments without departing from the broad general scope of the present disclosure. Accordingly, the present embodiments are to be considered in all respects as illustrative and not restrictive.

Description of Reference Numerals

[0200] 100 Filtration device, filter 102 Container 104 Inlet end 106 Outlet end 108 Inlet port 110 Outlet port 112 Flange 114 Opening 120 First filter unit 130 Second filter unit 200 Wall 202 Front wall 202-1 "Normal" state 202-2 Bulged state 204 Rear wall 206 Side end 208 Side end 210 Upper connecting part, seam 212 Lower connecting part, seam 212 Bottom seam 220 Spacer system 222 Separator 224 Separator body 226 Fence 228 Channel 230 Clip 240 Magnet system 242 First magnetic part 244 Second magnetic part 246 Metal structure 248 Metal structure 250 Brace 300 Chamber 310 Inlet chamber 320 Outlet chamber 330 Intermediate chamber 340 First front seam 342 First trough 350 First back seam 352 First peak 360 Second front seam 362 Second peak 370 Second back seam 370-1 First position 370-2 Second position 372 Second trough 400 System 500 Inlet conduit 510 Filling tube 520 Fluid source 520A First fluid source 520B Second fluid source 530 Inlet coupling 540 First inlet tube 550 Second inlet tube 560 Manifold 600 Outlet conduit 610 Drain tube 620 Vial 630 Outlet coupling

Claims

1. It is a filter, A container having an inlet port defined toward the inlet end of the container, an outlet port defined toward the outlet end of the container, and a flexible front wall and rear wall connecting the inlet end and the outlet end, A first filter unit and a second filter unit are spaced apart within the container, The first filter unit is coupled to the front and rear walls of the container and defines an inlet chamber that communicates with the inlet port. The second filter unit is coupled to the front and rear walls of the container and defines an outlet chamber that is in fluid communication with the outlet port. The first filter unit and the second filter unit define an intermediate chamber between the inlet chamber and the outlet chamber, the first filter unit is configured to filter the fluid flowing from the inlet chamber to the intermediate chamber, and the second filter unit is configured to filter the fluid flowing from the intermediate chamber to the outlet chamber, comprising the first filter unit and the second filter unit, The first filter unit is coupled to the front wall along the first front seam, and the second filter unit is coupled to the front wall along the second front seam. The first filter unit is coupled to the rear wall by a first back seam, and the second filter unit is coupled to the rear wall by a second back seam. The distance between the first front seam and the second front seam is smaller than the distance between the first back seam and the second back seam. A filter in which, when the fluid flows into the outlet chamber through the second filter unit, the outlet port and the second filter unit are configured to be spaced apart from each other.

2. The outlet port and the second filter unit are separated, and (i) the front wall and the first filter unit, (ii) The rear wall and the first filter unit, and (iii) The filter according to claim 1, further comprising a spacer system configured to separate at least one of the rear wall and the second filter unit.

3. The filter according to claim 1, wherein the first filter unit is connected at an acute angle to the front wall of the container and defines an inlet chamber trough.

4. The filter according to claim 1, wherein the second filter unit is connected at an acute angle to the rear wall of the container and defines an intermediate chamber trough.

5. The filter according to claim 1, wherein the first filter unit comprises a first filter mesh, and the second filter unit comprises a second filter mesh.

6. The filter according to claim 5, wherein the first filter mesh defines pores having an average pore diameter of 130 μm to 170 μm.

7. The filter according to claim 6, wherein the average pore size of the pores in the first filter mesh is 150 μm.

8. The filter according to claim 5, wherein the second filter mesh defines pores having an average pore diameter of 20 μm to 60 μm.

9. The filter according to claim 8, wherein the average pore size of the pores in the second filter mesh is 40 μm.

10. The filter according to claim 2, wherein the spacer system comprises a separator, a clip, a magnetic system, or a brace.

11. The filter according to claim 1, wherein the front wall is configured to bulge away from the second filter unit.

12. The filter according to claim 1, wherein the first filter unit and the second filter unit are angled relative to each other so as to be approximately V-shaped.

13. The filter according to claim 1, wherein the first filter unit and the second filter unit are angled relative to each other such that the cross-section of the intermediate chamber is generally triangular or trapezoidal.

14. The filter according to claim 1, wherein the container comprises a flexible bag.

15. The filter according to claim 1, wherein the first filter unit and the second filter unit are not parallel.

16. The filter according to claim 5, wherein when the fluid passes through the filter, at least one of the first filter mesh and the second filter mesh is spaced apart from the front wall or the rear wall.

17. The filter according to claim 1, wherein one or more or all of the container, the bag, or the filter unit are made of a DMSO-compliant plastic comprising one or more of polyethylene terephthalate (PET), high-density polyethylene (HDPE), low-density polyethylene (LDPE), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), thermoplastic elastomer (TPE), and / or polypropylene (PP).

18. A kit for filtering cells, A filter according to any one of claims 1 to 17, It is an inlet conduit, A first inlet tube and a second inlet tube are fluid-connected to the manifold, A filling tube is in fluid communication with the manifold and is adapted to receive fluid from at least one of the first inlet tube and the second inlet tube, The inlet conduit comprises an inlet coupling adapted to connect the filling tube to the inlet port of the filter and enable fluid communication with the inlet chamber, It is an outlet conduit, Drain tube and A kit comprising: an outlet conduit, an outlet coupling adapted to connect the drain tube to the outlet port of the filter and to enable fluid communication between the outlet chamber and the drain tube.

19. The kit according to claim 18, wherein the inlet tube and the filling tube form a Y-shape or a T-shape.

20. A method for filtering a fluid, comprising using a filter according to any one of claims 1 to 17, wherein the fluid is (i) a stem cell culture medium comprising cells, or (ii) a cell composition.

21. The method according to claim 20, comprising passing cultured cells through the filter to reduce visible particles and / or cell aggregates in order to produce a purified cell composition, wherein the first filter unit comprises a first filter mesh having an average pore size of 130 μm to 170 μm, and the second filter unit comprises a second filter mesh having an average pore size of 20 μm to 60 μm.

22. The method according to claim 21, wherein the cultured cells are provided in serum-free cell medium and / or the cells are cultured and grown, and / or the cells are mesenchymal progenitor cells or stem cells (MLPSCs).

23. The method according to claim 21, wherein, after passing the cells through the filter, the recovery rate of the viable cell concentration is (i) 60% to 100%, or (ii) 70% to 90%.

24. The purified cell composition contains D particles less than 150 μm, less than 100 μm, or less than 50 μm. ９０ The method according to claim 21, which demonstrates the present invention.

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