Container for containing degradable carrier

Abstract

To provide methods and containers for cultivating a biological agent.SOLUTION: A container 100 has an inner surface 114 comprising a fluoropolymer, and a degradable carrier stored in the container. The degradable carrier comprises a plurality of degradable microcarriers, each of the degradable microcarriers comprising a body formed from a hydrolyzable polymer, the body having a surface and a plurality of cell-capturing moieties attached to the surface. Alternatively, the degradable carrier comprises a coating disposed adjacently to at least a majority of the inner surface of the container, the coating comprising a body formed from a hydrolyzable polymer, the body having a surface and a plurality of cell-capturing moieties attached to the surface. The hydrolyzable polymer is one or more polymers selected from starch, polygalacturonic acid, poly(lactic-co-glycolic acid) (PLGA), polylactic acid (PLA), and polycaprolactone.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. 62 / 786,913, filed December 31, 2018. No. 6,313,535, filed on Oct. 13, 2003, which is hereby incorporated by reference in its entirety.

[0002] The present disclosure generally relates to a container (such as a bag) that contains a degradable carrier. The present disclosure provides a method for preparing a degradable carrier that includes a fluoropolymer and that includes a biological agent capture moiety. A container such as a bag for storing a cell, a cell isolation system including such a bag, and a cell isolation system including such a bag The present invention relates to a method for culturing biological agents using the same. [Background technology]

[0003] Cell culture and cell isolation are important processes in many applications, e.g., therapeutic Certain cells for use in applications (e.g., immunotherapy, regenerative medicine, etc.) are typically Cells, such as progenitor cells and mesenchymal stem cells, and Monocytes and other immune cells are present in the blood in relatively low concentrations and therefore typically They are isolated from blood and cultured in vitro. Similarly, neurons, cardiac myocytes, epithelial cells, and and other cells for regenerative medicine (e.g., bone repair, skin repair, pancreatic islet regeneration, etc.) It can be cultured in.

[0004] Fluoropolymer bags are commonly used for cell culture. Such bags are typically Typically, they are inexpensive, disposable, portable, and easy to use. The feed medium is mixed within the culture bag. As nutrients are consumed and cellular waste products accumulate, the feed medium is The medium must be removed and replenished; the cells can remain suspended in the bag. The desired cells can then be removed along with the depletion medium.

[0005] One conventional cell isolation process involves selectively immobilizing the desired cells on a solid support. For example, T cells, which play a central role in cell-mediated immunity, express T They can be distinguished from other lymphocytes by the presence of a T cell receptor (TCR). The solid support having a binding protein capable of binding to T cells in cell culture The captured cells are selectively captured while the undesired cells remain in suspension. Releasing the isolated cells can be accomplished, for example, by cleaving the anchor protein. This can potentially cause undesirable changes to the cells that are being treated.

[0006] Thus, there remains a need for cell culture and isolation articles that facilitate improved retention of desired cells. It is needed. Summary of the Invention

[0007] In one aspect, the present disclosure provides a container (e.g., in the form of a bag) having an exterior surface and an interior surface. The interior surface comprises a fluoropolymer, the container contains a degradable carrier, and the carrier comprises: A container is provided that contains a plurality of biological agent capture moieties.

[0008] In another aspect, the present disclosure provides a method for culturing a biological agent, the method comprising: providing a container having a biological agent therein; mixing the biological agent and the carrier in the first aqueous medium; and removing the first aqueous medium from the container. removing the first aqueous medium from the vessel; adding a second aqueous medium to the vessel; and and c) disintegrating the material.

[0009] Other aspects of the present invention will be apparent to those skilled in the art in view of the disclosure herein. cormorant. [Brief description of the drawings]

[0010] [Figure 1] 1A-1D are schematic top view (top) and cross-sectional view (bottom) of a bag according to one embodiment of the present disclosure.

[0011] [Diagram 2] FIG. 1 is a schematic top view of a bag according to certain embodiments of the present disclosure.

[0012] [Diagram 3] FIG. 1 is a schematic cross-sectional view of a bag according to an embodiment of the present disclosure.

[0013] [Figure 4] FIG. 1 is a schematic cross-sectional view of a bag according to an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present disclosure provides a degradable support having an inner surface that includes a fluoropolymer and that includes a biological agent capture moiety. The present disclosure relates to a container (e.g., a bag) for containing a body. The carrier can be physically separated from the suspension, and the degradation of the carrier does not adversely affect cell function. It is demonstrated that the suspension can be further resuspended without adverse effects.

[0015] The container of the present disclosure can be provided in a number of forms. One particularly convenient form is For example, a sheet or sheets of the fluoropolymer material described herein may be formed. Those skilled in the art will be familiar with bag structures such as those used in cell culture. Based on the description herein, conventional bag constructions may be used for the bags and methods of the present disclosure. Of course, those skilled in the art can adapt the container of the present disclosure to many other configurations. It will be appreciated that the formulation may be provided in a variety of formats, such as flasks, tubes, dishes, etc.

[0016] Thus, one aspect of the present disclosure is a container, for example in the form of a bag having an outer surface and an inner surface. and the inner surface comprises a fluoropolymer. One embodiment of such a bag is shown diagrammatically in FIG. The bag 100 of FIG. and a bag wall 110 having an inner surface 114, for transporting media (e.g., located at either end of the bag for adding or removing (e.g., feed medium or waste medium, respectively) The number and location of the ports are not particularly limited. Thus, those skilled in the art will appreciate that the present invention may be arranged for convenience of use or manufacture, for example. It will be appreciated that the bag 100 may be folded at its edges (e.g., For example, laser welding, corona discharge, radiation, heat or fusion lamination, etching, plasma treatment, wet Two fluoropolymer-containing sheets are attached together (by means of wetting, adhesives, or a combination thereof) A sheet (e.g., two sheets having a layer of fluorinated ethylene propylene sheet on their inner surfaces) The ports 130 and 140 may be products that couple the sealed compartment 12. It may be sealable to provide 0.

[0017] The walls of the container may be uniform in composition or may be made up of two or more different dots. The main layer may include, for example, two or more layers. By bonding polymer sheets together and then coating the bonded sheets, It is possible to provide an outer surface 112 that is different in composition from the inner surface 114. Similarly, two multi-layer systems can be provided. The grooves are joined together to provide an outer surface 112 that is different in composition from the inner surface 114. The multi-layer sheet can be made of both fluoropolymer and non-fluorinated polymer materials. In such a case, the inner surface of one or more of the multi-layer sheets may be formed with a ? The thickness of the container, the volume of the compartment, and the container and / or The shape of the compartment is not particularly limited, and may be determined for convenience of use or manufacture, and / or It can be selected to suit a particular application. For example, the wall thickness of the container can be 0.00 The volume of the compartment can range from 100 mL to 100 mL. 0L.

[0018] FIG. 2 shows some examples of culture bag configurations suitable for use with the bags and methods of the present disclosure. FIG. 1 shows an exemplary embodiment in which the bag 200a has only a single port 230a. Bag 200b provides access to compartment 220a. Bag 200b provides a longer path through the system. The ports 230b and 240 are in a so-called "serpentine" configuration that can provide a b is a serpentine-shaped section 220b formed by suitable welding of the sheets forming the bag. The bags 200c are connected by a meandering path formed by the above-mentioned. and a corresponding non-rectangular section 240c between ports 220c and 230c. .

[0019] One or more of the walls of the container may be porous, e.g., to accommodate the formation and growth of cells in cell culture. It is permeable to gases (e.g., O2, CO2) consumed, but not to liquids (e.g., water). ) to allow the cells in the bag to breathe. , allowing passive exchange of gas with the atmosphere across the vessel walls.

[0020] The containers of the present disclosure are desirably configured to be substantially free of contamination of the fluid within the container. Therefore, it is desirable that the inner surface of the container be made of a material that does not leach organic matter into the fluid. For example, in certain embodiments described elsewhere herein, the inner surface of the wall of the container , 0.1 mg / cm 2 Less than (e.g., 0.05 mg / cm 2 Less than or equal to 0.05 mg / cm 2 Polymers (e.g., fluorinated ethylene) with total organic carbon (TOC) in water of less than Such containers are made of, for example, fluoropolymers such as polyethylene terephthalate (PE) and polypropylene. The present invention is described in the published patent applications Nos. 2016 / 0178490 and 2016 / 0178491. each of which is incorporated herein by reference in its entirety. Based on the description herein, the present disclosure provides a method for the manufacture of such containers and methods. In particular, the container of the present disclosure has reactive functional groups on its inner surface. Although not required, in some cases, for example when the degradable carrier is a degradable coating, The degradable carrier is covalently attached to the interior surface of the container using reactive functional groups, as described below. It is possible.

[0021] The container contains a degradable carrier, the carrier comprising a plurality of biological agent capture moieties. When used, the biological agent capture moiety is a molecule to which a biological agent can selectively bind. The association between the biological agent and the respective capture moiety can be achieved by (e.g., centrifugation or sufficient to retain the biological agent on the carrier during the course of the carrier (e.g., during filtration or filtration), and is covalently bonded In certain embodiments described elsewhere herein, The biological agent capture moiety comprises one or more aptamers. In some embodiments, the one or more aptamers may be peptides, polypeptides, oligonucleotides, or the like. and polynucleotides (i.e., DNA and RNA). In certain embodiments described elsewhere herein, the biological agent capture moiety is a randomly synthesized The aptamers include oligonucleotides selected from a pool of synthetically generated sequences.

[0022] In certain embodiments described elsewhere herein, the biologic agent capture moiety is a DNA aptamers having at least two sites to which a biologic agent can selectively associate - Patents.com For example, in certain such embodiments, a nucleotide sequence selective for a biological agent is At least 5, or at least 10, or at least 25 of the sequence of has at least 50, or at least 75, or at least 100 instances of A is present in the aptamer sequence.

[0023] In certain embodiments described elsewhere herein, the biological agent capture moiety is a poly Contains multiple aptamers (e.g., oligonucleotide aptamers) bound to a peptide For example, in certain embodiments described elsewhere herein, the biological agent capture moiety The antibody comprises a plurality of biotinylated aptamers bound to an avidin protein. In certain embodiments, the biotinylated aptamer comprises an oligonucleotide or a polypeptide. In certain such embodiments, the biological agent capture moiety comprises at least 10 or At least 25, or at least 50, or at least 75, or at least At least 100, or at least 200, or at least 300, or at least 400, or at least 500, or at least 600, or at least 7 00, or at least 800, or at least 900, or at least 10 00 aptamers (e.g., biotinylated aptamers bound to avidin protein) In certain such embodiments, the biological agent capture moiety comprises 10 to 1000, or is 10~900, or 10~800, or 10~700, or 10~600, or is 10~500, or 10~400, or 10~300, or 10~200, or is 25~1000, or 50~1000, or 100~1000, or 200~1 000, or 300-1000, or 400-1000, or 500-1000, or 600-1000, or 700-1000, or 800-1000, or 1 0~400, or 100~500, or 200~600, or 300~700, or or any number within the range of 400-800, 500-900, or 600-1000 The aptamer includes an aptamer (e.g., a biotinylated aptamer bound to an avidin protein). .

[0024] In certain embodiments described elsewhere herein, the biological agent is an inorganic species and For example, in certain such embodiments, the biological agent is selected from a gold In other such embodiments, the biological agent is a vitamin, a hormone, or a peptide. In certain embodiments as otherwise described herein, The agent may be, for example, a protein, an enzyme, or a nucleic acid (i.e., DNA or RNA). In certain embodiments described elsewhere herein, the biological agent is , e.g., organelles (e.g., nuclei, ribosomes, mitochondria, vacuoles, rough endoplasmic reticulum, Complex biological tissues such as smooth endoplasmic reticulum, Golgi apparatus, lysosomes, centrosomes, vesicles, membranes, or cell fragments. In certain embodiments described elsewhere herein, the biological agent is a bacterial Or a group of cells.

[0025] In certain embodiments described elsewhere herein, the biological agent is a cell (e.g., For example, certain embodiments described elsewhere herein may be used. In certain such embodiments, the cells are blood cells, stem cells, or immune cells. In certain such embodiments, the cells are leukocytes. In certain such embodiments, the cells are monocytes. In such embodiments, the cells are stem cells, e.g., mesenchymal stem cells or progenitor cells. In certain embodiments described elsewhere herein, the cells are T cells (e.g., regulatory The biological agent may be a regulatory T cell, an endothelial progenitor cell, or a natural killer cell. The agent, in certain embodiments, is selected for retention in cell culture (i.e., positive selection). Alternatively, in another embodiment, the desired biological agent may be captured in the cell culture. (i.e., negative selection) You will understand that it could be.

[0026] For example, in certain embodiments described elsewhere herein, the biological agent capture moiety one or more antibodies capable of selectively associating with the CD31 marker of endothelial progenitor cells In another example, in certain embodiments described elsewhere herein, The biological drug capture moiety binds to one of the regulatory T cells (e.g., CD4, CD25, CD27). or a plurality of markers. In yet another example, certain embodiments described elsewhere herein include a biological agent capture The capture moiety may be one or more of the following: The aptamer comprises one or more aptamers capable of selectively associating with a marker of a certain In such embodiments, the biological agent capture moiety comprises multiple apeptides attached to the polypeptide. aptamers (e.g., oligonucleotide aptamers).

[0027] As used herein, a degradable carrier is a carrier that, in its undegraded state, is capable of trapping biological agents. A material having a surface to which a capture moiety can be attached (e.g., via a covalent linker). The carrier may be, for example, a material suitable for culturing or isolating a biological agent in the presence of an aqueous medium. The carrier may remain undegraded for at least a period of time, or even indefinitely. or may be partially or completely dissolved and may be biologically active in the decomposed state. It can be separated from the drug.

[0028] In certain embodiments described elsewhere herein, the biological agent capture moiety is For example, the surface of the degradable carrier may be covalently bonded to a functional group on the surface of the degradable carrier. group, hydroxyl group, aldehyde group, carbonyl group, amine group, imine group, amide group, ester groups, anhydride groups, thiol groups, disulfides, phenols, guanidines, thioethers The compound may be functionalized with an indole, an imidazole or a diazonium group. These "functional groups" are identified as groups to which biological agent capture moieties are covalently attached. For example, it will be appreciated that the "amine" functional group may be substituted with a carboxyl-containing biological agent. The compound can be linked to the biological agent capture moiety to form a carboxamide bond to the biological agent capture moiety. can.

[0029] In certain such embodiments, the biological agent capture moiety comprises multiple The protein comprises an aptamer (e.g., an oligonucleotide aptamer) of The peptide bonds are attached to the carboxyl groups on the surface of the carrier. In this form, the biological agent capture moiety is attached to the aldehyde groups on the surface of the degradable carrier via an imine bond. The aptamer comprises an amine-terminated aptamer (eg, a DNA aptamer) bound via a cation bond.

[0030] The number of biological agent entrapment moieties per unit of degradable carrier surface (i.e., entrapment moiety density) However, those skilled in the art will appreciate that the density of the capture moieties is typically determined by the number of units on the surface of the degradable carrier. The number of biological agents (e.g., cells) that can be captured per unit time (i.e., the captured agent density) It will be appreciated that the present invention may be implemented in a variety of ways, including, for example, in which the optical axis is a sine wave (S1) or a sine wave (S2). The capture moiety density of the degradable carrier is greater than the capture agent density of the degradable carrier.

[0031] In certain embodiments described elsewhere herein, the degradable support comprises a plurality of degradable microparticles. The microcarriers include small carriers (e.g., polymeric microcarriers) having attached thereto biological agent capture moieties. For example, the bag 300 of FIG. 310 (having an outer surface 312 and an inner surface 314) and housing a plurality of degradable microcarriers 350. In a particular embodiment, the average size of the microcarriers is between 0.5 μm and 1000μm, or 0.5μm to 900μm, or 0.5μm to 800μm, or 0.5μm~700μm, or 0.5μm~600μm, or 0.5μm~500μm m, or 0.5 μm to 400 μm, or 0.5 μm to 300 μm, or 0.5 μm ~250μm, or 1μm~1000μm, or 5μm~1000μm, or 10 μm~1000μm, or 25μm~1000μm, or 50μm~1000μm, Or 100μm~1000μm, 200μm~1000μm, or 300μm~10 00μm, or 400μm to 1000μm, or 500μm to 1000μm, or 10μm~750μm, or 10μm~500μm, or 20μm~400μm, or Or in the range of 50 μm to 300 μm.

[0032] In certain embodiments described elsewhere herein, the degradable carrier is a small amount of the inner surface of the container. at least a portion (e.g., at least a majority, or even all) of Degradable coatings (e.g., polymeric coatings) are included. In this embodiment, the degradable coating is covalently bonded to the interior surface of the container. For example, the bag of FIG. 400 includes a bag wall 410 (having an outer surface 412 and an inner surface 414) surrounding an enclosed compartment 420. ) and a degradable coating disposed adjacent (e.g., covalently bonded to) the inner surface 414. In certain such embodiments, the degradable core comprises a sealed compartment 420 that contains a degradable core 460. The coating may be from 10 nm (e.g., about a monolayer) to 10 mm, or from 10 nm to 1 mm, or is 10nm~500μm, or 10nm~100μm, or 10nm 1μm, or is 1μm~10mm, or 100μm~10mm, or 500μm~10mm, or In certain such embodiments, the degradable core has a thickness in the range of 1 mm to 10 mm. The coating is applied to the functional groups on the interior surface of the container (e.g., the functional groups of the fluoropolymer that comprises the interior surface). In certain embodiments, the functional group is a carboxyl group, a hydroxyl group, or a hydroxyl group. Silyl group, aldehyde group, carbonyl group, amine group, imine group, amide group, ester group, no Aqueous groups, thiol groups, disulfides, phenols, guanidines, thioethers, indomethacin, The substituent is an alkyl, imidazole or diazonium group.

[0033] In certain embodiments described elsewhere herein, the degradable carrier comprises a cleavable polymer. For example, in certain such embodiments, the degradable support may be an enzyme-cleavable, e.g., polysaccharide. In certain such embodiments, the polysaccharide comprises a starch (e.g., α-glycosylating polymer). In other such embodiments, the polysaccharide is a polysaccharide. Lacturonic acid (e.g., cleavable by pectinase). In certain such embodiments, the degradable carrier may be, for example, poly(lactic-co-glycolic acid). Hydrolyzable poly(propylene glycol arylene glycol) (PLGA), polylactic acid (PLA), or polycaprolactone In certain such embodiments, polymer hydrolysis occurs at a pH below 6 or In certain such embodiments, polymer hydrolysis is accelerated at a pH above 8. The reaction is accelerated in the presence of a hydrolysis catalyst, for example an amine.

[0034] In certain embodiments described elsewhere herein, the degradable carrier is, for example, an aqueous medium. A cross-linked polysaccharide having a sufficient number of cross-links to provide a carrier that can remain undegraded in the body. In certain such embodiments, the degradable carrier comprises an enzyme below a critical level. A sufficient number of crosslinks (e.g., Enzyme-cleavable polymers (e.g., polysaccharides) having glycerol diether crosslinks In other such embodiments, the degradable support has less than a critical level of hydrolysis catalyst. Carriers that can remain undegraded in aqueous media or in aqueous media having a pH outside the critical range A hydrolyzable polymer (e.g., PLGA, PLA) having a sufficient number of crosslinks to provide In another example, the specific polymers described in other formats herein include polycaprolactones. In certain embodiments, the degradable carrier comprises an insoluble crosslinked network of otherwise soluble polymers. For example, in certain such embodiments, a thioether cross-linked polymer network is Degradable carriers, including gels (e.g., hydrogels), have less than a critical level of glutathione. The compound may remain undegraded in aqueous media.

[0035] In certain embodiments, the cross-linked polymer of the degradable carrier comprises a cross-linking agent that prevents enzymatic cleavage of the polymer. For example, in certain such embodiments, Ca 2+ Multi-crosslinked Degradable carriers containing sugars (e.g., polygalacturonic acid) can retain Ca below a critical level. 2+ Angry In an aqueous medium with a thickening agent (e.g., EDTA), i.e., when the medium exceeds a critical level Even if the protein contains a cleavage enzyme (e.g., pectinase) that can cause digestion, it may remain undigested.

[0036] In certain embodiments described elsewhere herein, the degradable carrier is (e.g., 45 decomposable upon irradiation (with radiation having a wavelength of less than 420 nm or less than 420 nm) or accelerates the degradation of the degradable carrier. In an embodiment, the degradable carrier degrades when exposed to elevated temperatures (e.g., above 37° C., or above 40° C.). It is degradable or the degradation of a degradable carrier is accelerated.

[0037] In certain embodiments described elsewhere herein, the interior surface of the container is made of polytetrafluoroethylene. Perfluoroethylene (PTFE), Perfluoroalkoxy (PFA), Ethylene tetrafluoroethylene Polyethylene (ETFE), Polyvinylidene fluoride (PVDF), Polychlorotrifluoro Ethylene (PCTFE), Ethylene chlorotrifluoroethylene (ECTFE), Fluorine Fluorinated ethylene propylene (FEP), ethylene fluorinated ethylene propylene (EFEP), Perfluoropolyether (PFPE), Modified polytetrafluoroethylene (TFM), Fluoropolyethers selected from polyvinyl fluoride (PVF), or any mixture thereof. For example, in certain embodiments described elsewhere herein, the inner surface of the container includes fluorinated ethylene propylene. Certain embodiments as otherwise described herein In this case, the inner surface of the container is made of a fluoropolymer (e.g., fluorinated ethylene propylene). The specific polymers described elsewhere herein may be either fluoropolymers or fluoropolymers. In an embodiment, the material comprising the interior surface of the bag has a thickness of at least 0.0003 inches, at least 0.0004 inches, at least 0.0005 inches, at least 0.0006 inches, It has a thickness of at least 0.001 inches, or at least 0.10 inches. For example, In certain such embodiments, the material of the interior surface of the container is between 0.0003 inches and 0.2 inches. inches, or 0.0003 inches to 0.1 inches, or 0.0005 inches to 0.08 inches inch, or 0.001 inch to 0.07 inch, or 0.001 inch to 0.05 inch, or 0.001 inch to 0.03 inch, or 0.001 inch to 0.01 8 inches, or 0.001 inches to 0.016 inches, or 0.001 inches to 0. 014 inches, or has a thickness within the range of 0.001 inches to 0.012 inches.

[0038] In certain embodiments described elsewhere herein, the material that makes up the wall of the container is The inner surface of the fluoropolymer layer is coated with another polymer material (fluoropolymer or otherwise). In this embodiment, the material of the exterior of the container is at least 0.0005 inches thick, or at least At least 0.001 inches, or at least 0.005 inches, or at least 0.007 inches 5 inches, or at least 0.01 inches, or at least 0.02 inches, or At least 0.03 inches, or at least 0.04 inches, or at least 0.0 5 inches, or at least 0.06 inches, or at least 0.07 inches, or At least 0.08 inches, or at least 0.09 inches, or at least 0.1 inch, or at least 0.11 inches thick. For example, certain such In an embodiment, the material of the exterior of the container is between 0.0005 inches and 0.2 inches, or 0.00 5" to 0.18", or 0.01" to 0.16", or 0.01" inch to 0.14 inch, or 0.01 inch to 0.12 inch, or 0.06 inch ~0.13 inches, or has a thickness within the range of 0.09 inches to 0.126 inches.

[0039] In certain embodiments, the degradable carrier comprises a plurality of microcarriers, and the interior surface of the container is The biological agent capture moieties may further comprise a plurality of biological agent capture moieties as described in any other manner herein. In certain embodiments, the biological agent capture moieties attached to the interior surface of the container are biocompatible with the degradable carrier. In another embodiment, the biological agent capture moiety attached to the interior surface of the container is the same as the biological agent capture moiety. The capture moiety is selective for a biological agent other than the agent for which the biological agent capture moiety of the degradable carrier is selective. Advantageously, the inventors have demonstrated that such containers are effective in capturing biological agents. It can improve the efficiency of the purification or facilitate the isolation and separation of two different biological agents. It was determined that this could be done.

[0040] In certain embodiments described elsewhere herein, the exterior surface of the container is made of a fluoropolymer. For example, in certain such embodiments, the material of the exterior surface of the container is a thermally Thermoplastic polymers, thermoplastic elastomers, silicones, rubbers, or any combination thereof Alternatively, the exterior surface of the container may include any of the specific embodiments described elsewhere herein. For example, the inner surface of the fluoropolymer may include a fluoropolymer. In such an embodiment, the interior and exterior materials (i.e., the container walls) are fluoropolyethersulfones. The polymer may consist essentially of a fluoropolymer (e.g., fluorinated ethylene propylene) or may be a fluoropolymer. It's Rimmer.

[0041] In certain embodiments described elsewhere herein, the container contains a biological agent. In certain such embodiments, the biological agent is attached to the carrier via a biological agent capture moiety. For example, in certain such embodiments, the biological agent is incorporated into a degradable microcarrier or or cells associated with the carrier via aptamers bound to the surface of the degradable coating. In certain embodiments, the interior surface of the container comprises a plurality of biological agent capture moieties, at least For example, in certain such embodiments, The biological agent is a cell, an aptamer bound to the surface of a degradable microcarrier, or to the inner surface of a vessel. In certain such embodiments, the degradable The biological agent-entrapping portion of the microcarriers is the same as the biological agent-entrapping portion of the interior surface of the container.

[0042] In certain embodiments described elsewhere herein, the container further comprises an aqueous medium. The aqueous medium may be, for example, a cellular solution containing a biological agent and one or more off-target agents. As used herein, an off-target agent may be, for example, a biological Cells, cell fragments, proteins, vitamins, hormones, peptides, and gold, other than biological drugs Materials other than the biological agent (i.e., materials that are not captured by the biological agent capture moiety) that contain metal ions (It is possible to grasp this.)

[0043] In certain embodiments described elsewhere herein, the container may contain off-target agents (e.g. (e.g., cells other than biological agents) and that no more than 20% of the off-target agents are delivered via degradable carriers or For example, in certain such embodiments, less than 17.5% , or less than 15%, or less than 12.5%, or less than 10%, or less than 7.5%, Or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% The off-target agent is attached to the interior surface of the degradable support or container.

[0044] Another aspect of the present disclosure is a cell isolation system comprising a vessel as described elsewhere herein. For example, in certain embodiments described elsewhere herein, a cell isolation system The container described herein contains an aqueous medium, and the aqueous medium and the carrier are mixed together. In another example, the locker may include a locker configured to fit into the locking mechanism as described in other manners herein. In certain embodiments, the cell isolation system comprises a cell isolation system as described elsewhere herein that contains an aqueous medium. Separating the carrier (e.g., comprising a plurality of degradable microcarriers) from the vessel in which it is placed and from the aqueous medium. In yet another example, the centrifuge includes a centrifuge configured to In certain embodiments described herein, the cell isolation system comprises a cell suspension containing an aqueous medium. and a container for separating a carrier (e.g., comprising a plurality of degradable microcarriers) from an aqueous medium. and a filter in fluid communication with the outlet of the vessel configured to The aqueous medium in the vessel of the cell isolation system may be used to isolate off-label substances such as cells other than biological agents. -Contains a target agent.

[0045] Advantageously, the inventors have demonstrated that degradation of the carriers described herein does not adversely affect cellular function. The captured (e.g., isolated) biological agent may be transferred to an aqueous medium (e.g., It was determined that the cells could be released into the environment (if necessary, they should be stored or further cultured). Desirably, the inner surface fluoropolymer is free of any material other than the biological agent (i.e., off-agent). This prevents adhesion of the target agent to the container and increases the purity of the isolated biological agent in the container. do.

[0046] Accordingly, another aspect of the present disclosure is a method for culturing a biological agent, comprising: providing a container as described elsewhere herein containing a therapeutic agent; and mixing the biological agent and the carrier in a first aqueous medium contained therein. The method includes removing a first aqueous medium from a container and introducing a second aqueous medium into the container. The method further includes the steps of adding and decomposing the carrier.

[0047] In certain embodiments described elsewhere herein, the container contains a blood sample containing a biological agent. For example, in certain such embodiments, the container In certain embodiments, the container comprises one or more For example, in certain such embodiments, the container further comprises a plurality of off-target agents. The method involves the administration of biological agents (e.g., monocytes, stem cells, T cells, endothelial progenitor cells or natural killer cells). -targeted cells) and one or more off-target agents (e.g., cells other than the biological agent). The sample includes a sample (e.g., a whole blood sample).

[0048] In certain embodiments described elsewhere herein, the first aqueous medium is a cell culture medium. In certain such embodiments, the biological agent is a cell, and the biological agent is a The agent is cultured in a vessel (i.e., the agent grows or expands in the first aqueous medium). Of course, one or more of the above may be used in combination with an aqueous medium (e.g., a whole blood sample contained in a container). The off-target agents may also swell in the first aqueous medium. In the particular embodiment described, the biological agent and the carrier are mixed on a rocker. The mixing is carried out for a period of time sufficient to allow association of the biological agent capture moiety with the biological agent. In certain embodiments described elsewhere herein, the biological agent is a cell and the mixture is , for a time and at a temperature sufficient to allow proliferation of the biological agent. The mixing is carried out in a rocker.

[0049] In certain embodiments described elsewhere herein, the carrier comprises a plurality of degradable microcarriers. The step of removing the first aqueous medium includes settling the carrier and removing the first aqueous medium from the vessel. For example, in certain such embodiments, the first aqueous medium is decanted. The step of removing the bodies may include centrifuging the microcarriers and removing the first and decanting the aqueous medium. In an embodiment, the first aqueous medium is passed through a filter configured to retain the microcarriers. For example, in certain such embodiments, the first aqueous medium is The degradable microcarriers are then removed from the container through a filter having a smaller pore size than the undegraded microcarriers. Removed.

[0050] In another embodiment, the carrier comprises a degradable coating disposed adjacent to the interior surface of the container. and removing the first aqueous medium includes decanting the first aqueous medium from the container. i.e., without sedimentation or filtration).

[0051] Of course, one of ordinary skill in the art will understand that a small amount of the first aqueous medium can be retained in the container. You will understand.

[0052] After removal of the first aqueous medium, the container as otherwise described herein may be filled with, for example, a biological agent. The composition may contain cells that contain the agent and optionally one or more off-target agents. Desirably, the majority of the cells remaining in the container are degraded to the degradable carrier and optionally The container may include a biological agent attached to an interior surface via a biological agent capture moiety. For example, in certain embodiments described elsewhere herein, the biological agent is a cell. and at least 80% of the cells in the container after removing the first aqueous medium, or at least 82.5%, or at least 85%, or at least 87.5%, or at least 90%, or at least 92.5%, or at least 95%, or at least 97% .5%, or at least 98%, or about 98.5%, or at least 99% biological It is a therapeutic drug.

[0053] In certain embodiments described elsewhere herein, the step of degrading the support comprises degrading the support. The method includes subjecting the body to a pH that is different from the pH of the first aqueous medium. In certain embodiments described otherwise, the step of degrading the support comprises degrading the support to less than 6 In certain such embodiments, the container includes a pH of 8 or greater. The second aqueous medium used has a pH of less than 6 or greater than 8. The pH of the second aqueous medium is adjusted after adding the second aqueous medium to the container. In certain embodiments described elsewhere herein, the step of decomposing the support comprises dehydrating the support. The method includes contacting the ester with a decomposition catalyst (e.g., an amine catalyst). In this embodiment, the second aqueous medium added to the vessel contains a hydrolysis catalyst. In an embodiment, the hydrolysis catalyst is added to the vessel after the second aqueous medium is added. In an embodiment, the pH of the second aqueous medium is adjusted to favor the hydrolysis reaction catalyzed by the hydrolysis catalyst. Certain embodiments described elsewhere herein In the method, the step of decomposing the carrier comprises contacting the carrier with an enzyme, or an enzyme and a chelating agent. In certain such embodiments, the second aqueous medium added to the vessel comprises In other such embodiments, the enzyme is added after the second aqueous medium is added. In certain embodiments described elsewhere herein, the carrier is degraded by adding The step of subjecting the carrier to an elevated temperature (e.g., a temperature above 37° C. or above 40° C.) In certain embodiments described elsewhere herein, the carrier is decomposed. The step of irradiating light includes irradiating light having a wavelength of less than 450 nm or less than 420 nm. The method includes irradiating the carrier (with radiation).

[0054] Advantageously, we degrade the carrier (e.g., fragment, partially dissolve, etc. or dissolving the biological agent, thereby causing adverse effects on the biological agent. They found that the captured biological agent can be separated from the carrier without causing any damage to the carrier. In certain embodiments described elsewhere herein, the method further comprises administering the biological agent to a second The method further comprises culturing the bacteria in an aqueous medium (i.e., containing the degraded carrier). In such an embodiment, the second aqueous medium is a cell culture medium. In certain embodiments described above, the method further comprises precipitating the biological agent and then adding a second aqueous solution to the biological agent. The decomposed carrier is removed by removing the aqueous medium and adding a third aqueous medium to the vessel. In certain such embodiments, the third aqueous medium further comprises: A culture medium or a cell storage medium.

[0055] Further aspects of the present disclosure include any number and any method that is not technically or logically inconsistent. The present invention is provided by the following embodiments, which can be combined in any combination: [Embodiment 1] A container (e.g., in the form of a bag) having an exterior and an interior surface, the interior surface being made of fluoropolyether. The degradable carrier is contained in a container. A container, wherein the carrier comprises a plurality of biological agent capture moieties. [Embodiment 2] The degradable carrier comprises a plurality of degradable microcarriers, each microcarrier having a plurality of biological agent capture moieties. 2. The container of embodiment 1, comprising: [Embodiment 3] The average diameter of the microcarriers is about 0.5 μm to about 1000 μm (for example, about 5 μm to about 750 μm). m, or about 10 μm to about 500 μm, or about 25 μm to about 300 μm 3. The container of embodiment 2. [Embodiment 4] 4. A container according to embodiment 2 or 3, wherein the interior surface of the container comprises a plurality of biological agent capture moieties. . [Embodiment 5] The degradable carrier is adjacent to at least a portion (e.g., at least a majority) of the inner surface of the container. 2. The container of embodiment 1, comprising a degradable coating disposed thereon. [Embodiment 6] Any of embodiments 1 to 5, wherein the biological agent capture moiety comprises one or more aptamers. or one of the containers described in. [Embodiment 7] 7. The container of any one of embodiments 1 to 6, wherein the biological agent is a cell. [Embodiment 8] 8. The container of embodiment 7, wherein the cells are blood cells or immune cells. [Embodiment 9] The cell may be a monocyte, a stem or progenitor cell (e.g., a mesenchymal stem cell), a T cell (e.g., Regulatory T cells), endothelial progenitor cells or natural killer cells, container. [Embodiment 10] 10. The container of any one of embodiments 1-9, wherein the degradable carrier comprises a polymer. [Embodiment 11]

[0023] Embodiment 1, in which the degradable carrier comprises a polysaccharide (e.g., starch or polygalacturonic acid). Container as described in 0. [Embodiment 12] Degradable carriers include poly(lactic-co-glycolic acid) (PLGA) and polylactic acid (PLA). and polycaprolactone. Container as described in 0. [Embodiment 13] The polymer is cross-linked (e.g., thioether cross-linked, Ca 2+ Crosslinking, or glycerol diene 13. The container according to any one of embodiments 10 to 12, wherein the container is ether-crosslinked. [Embodiment 14]

[0023] Embodiments 1-1, wherein the carrier is degradable in water at a pH below 6 or above 8. 3. A container according to any one of claims 1 to 3. [Embodiment 15] The support is decomposable in water when contacted with a hydrolysis catalyst (e.g., an amine catalyst). A container according to any one of embodiments 1 to 13. [Embodiment 16] The carrier is degradable in water when contacted with an enzyme, or an enzyme and a chelating agent. A container according to any one of embodiments 1 to 13. [Embodiment 17] 17. The container of embodiment 16, wherein the enzyme is an amylase or a pectinase. [Embodiment 18] When the carrier is exposed to high temperatures (e.g., temperatures above 37°C or above 40°C), 14. The container according to any one of embodiments 1 to 13, which is decomposable in water. [Embodiment 19] The carrier is irradiated with radiation having a wavelength of less than 450 nm, or less than 420 nm, for example. The container according to any one of embodiments 1 to 13, which is decomposable in water upon irradiation. [Embodiment 20] Fluoropolymers are polytetrafluoroethylene (PTFE), perfluoroalkane, Polyvinylidene fluoride (PFA), Ethylene tetrafluoroethylene (ETFE), Polyvinylidene fluoride (PVDF), Polychlorotrifluoroethylene (PCTFE), Ethylene Chlorotrifluoroethylene Fluoroethylene (ECTFE), Fluorinated Ethylene Propylene (FEP), Ethylene Fluoride Perfluoroethylene propylene (EFEP), perfluoropolyether (PFPE), modified poly Polytetrafluoroethylene (TFM), polyvinyl fluoride (PVF), or any of the above 20. The container according to any one of embodiments 1 to 19, which is a mixture of any of the above. [Embodiment 21] Embodiments 1-19, wherein the fluoropolymer is fluorinated ethylene propylene (FEP). 3. A container according to any one of the preceding claims. [Embodiment 22] The degradable carrier is bound to a biological agent, and the biological agent is released via the biological agent capture moiety. 22. The container according to any one of embodiments 1 to 21, wherein the container is bound to a carrier. [Embodiment 23] 23. A container according to any one of embodiments 1 to 22, containing an aqueous medium. [Embodiment 24] 24. The container of embodiment 23, containing an off-target agent. [Embodiment 25] 25. The container of embodiment 24, wherein the off-target agent is a cell other than a biological agent. [Embodiment 26] Less than 20% (e.g., less than 15%, less than 10%, less than 5%, or 26. The container of embodiment 24 or 25, wherein less than 1% of the carrier or container is attached to the inner surface of the carrier or container. vessel. [Embodiment 27] A container according to any one of embodiments 23 to 25 and a container for mixing a carrier and an aqueous medium. A cell isolation system comprising: a locker configured as above. [Embodiment 28] A container according to any one of embodiments 23 to 25 and a container for separating a carrier from an aqueous medium. and a centrifuge configured to: [Embodiment 29] A container according to any one of embodiments 23 to 25 and a filter in fluid communication with the outlet of the container. and a filter configured to separate the carrier from the aqueous medium. system. [Embodiment 30] 1. A method for culturing a biological agent, comprising: A container according to any one of the first to twenty-first embodiments, containing a biological agent. Providing a container; mixing a biological agent and a carrier in a first aqueous medium contained in a container; removing the first aqueous medium from the vessel; adding a second aqueous medium to the vessel; Decomposing the support; The method includes: [Embodiment 31] The carrier comprises a plurality of degradable microcarriers, and the step of removing the first aqueous medium comprises removing the microcarriers and decanting the first aqueous medium from the container. 31. The method of embodiment 30, comprising: [Embodiment 32] The carrier comprises a plurality of degradable microcarriers, and the first aqueous medium is configured to retain the microcarriers. 31. The method of embodiment 30, wherein the liquid is removed from the container through a filter formed therein. [Embodiment 33] The carrier includes a degradable coating disposed adjacent to an inner surface of the container, and a first aqueous medium In one embodiment, the step of removing comprises decanting the first aqueous medium from the container. 31. The method according to claim 30. [Embodiment 34] Biological agents are cells; At least 80% of the cells in the container after removing the first aqueous medium (e.g., at least 85%, or at least 90%, or at least 95%, or at least 98%, or at least 99%) is a biological agent; The method according to any one of embodiments 30 to 34. [Embodiment 35] The step of degrading the carrier may include subjecting the carrier to a pH of less than about 6 or less than about 8. The method of any one of embodiments 30 to 34, comprising: [Embodiment 36] The step of decomposing the support includes contacting the support with a hydrolysis catalyst (e.g., an amine catalyst). The method according to any one of embodiments 30 to 34, comprising the step of: [Embodiment 37] The step of degrading the carrier comprises contacting the carrier with an enzyme and optionally a chelating agent. The method of any one of embodiments 30 to 34, comprising: [Embodiment 38] The step of decomposing the carrier may include subjecting the carrier to high temperatures (e.g., above 37° C. or below 40° C.). The method of any one of embodiments 30 to 34, comprising exposing the composition to a temperature exceeding 1000° C. Law. [Embodiment 39] The step of decomposing the carrier may include, for example, using a wavelength of less than 450 nm or less than 420 nm. 35. Any of embodiments 30 to 34, comprising irradiating the carrier (with radiation having a long wavelength). The method described in one. [Embodiment 40] Any of embodiments 30 to 39, further comprising removing the degraded carrier from the container. or one of the methods described above. [Embodiment 41] 41. The container or method of any one of claims 1 to 40, wherein the container is in the form of a bag.

[0056] The details given herein are given by way of example and for purposes of discussing preferred embodiments of the invention. Only the above-mentioned embodiments are presented and are intended to provide the most useful understanding of the principles and conceptual aspects of various embodiments of the present invention. The present disclosure is presented to provide an explanation that is believed to be readily understandable to those skilled in the art. In this regard, No attempt is made to show structural details of the invention in more detail than is necessary for the fundamental understanding of the invention. First, the drawings and / or examples will illustrate how some forms of the invention may be It will be apparent to one skilled in the art how the disclosed processes and devices can be implemented. Before describing the present invention, it should be understood that the aspects described herein may be implemented in a specific embodiment, device, or configuration. It is understood that the present invention is not limited to the above and as such may, of course, vary. Further, the terminology used herein is not intended to describe particular embodiments. It is understood that no limitation is intended unless specifically defined herein. I want to be.

[0057] Terms used in the context of describing the present invention (especially in the context of the claims below) "A," "an," "the," and similar referents are used herein unless otherwise indicated. Unless otherwise specified or clearly contradicted by the context, both the singular and the plural are intended to be covered. The recitation of a range of values ​​herein should be construed as merely including within that range. The term "value" is intended to serve as a shorthand for referring individually to each separate value in the specification. Unless otherwise indicated, each individual value is incorporated herein as if it were individually set forth herein. As used herein, certain limits may be used to express a range of values, including but not limited to, from "about" one particular value and / or from "about" another particular value. A range can be expressed up to "about." When expressing such a range, another embodiment is as follows: Including from one particular value and / or up to another particular value. Similarly, values ​​may be rounded to the nearest whole number. When expressed, the use of the antecedent "about" may be used to indicate that the particular value constitutes another aspect. It will be understood that the endpoints of each of the ranges may be expressed relative to the other endpoint and independently of the other endpoint. It will be understood that both are important.

[0058] All methods described herein are intended to be illustrative, unless otherwise indicated herein or by context. Any suitable order of steps may be performed unless clearly contradicted. Any and all examples provided in the specification, or the use of exemplary language (e.g., "etc.") The use is merely intended to make the invention more readily understandable and is not intended to be a substitute for the invention as otherwise claimed. No language in this specification is intended to be limiting in scope. It should not be construed as indicating non-claimed elements.

[0059] Unless the context clearly requires otherwise, throughout this specification and claims In this regard, words such as "comprise" and "comprising" are used to mean exclusive or exhaustive. should be interpreted in an inclusive sense, not in a restrictive sense, i.e. "including, but not limited to, The term "a," "the ... In addition, as used in this application, "the present specification", "the present application ... The words "greater than," "less than," and words of similar meaning may be used in any particular portion of this application. "Patent application 10 / 113, 1997-10 / 13" refers to this application in its entirety.

[0060] As will be appreciated by those skilled in the art, each embodiment disclosed herein may be The invention relates to a method for producing a process for the production of ... As used herein, the transition word "comprises" "se" or "comprises" means an indefinite element, step, or even a major quantity. "Consists of" means to include, but is not limited to, a component, or ingredient. The transitional phrase "excludes any element, step, ingredient, or component not specified." The transition term "consisting essentially of" extends the scope of the embodiment to the specified elements, steps, and / or components. The present invention is limited to those groups, components or constituents that do not have a significant effect on the embodiment.

[0061] Unless otherwise specified, the amounts, molecular weights, and other information used in the specification and claims are All numbers expressing properties, reaction conditions, etc. are preceded in all cases by the word "about." Accordingly, unless otherwise indicated in the specification and accompanying drawings, The numerical parameters set forth in the following claims are intended to represent the desired characteristics sought to be obtained by the present invention. These are approximations that may vary depending on the nature of the material used. Without limiting the application of the doctrine of equivalents, each numerical parameter should at least represent the They should be interpreted taking into account significant digits and applying ordinary rounding techniques. When necessary, the term "about" is used in connection with a stated numerical value or range. has the meaning that would reasonably be given to it by a person of ordinary skill in the art when Within a range of degrees, it indicates something more or something less than the stated value or range.

[0062] Although the numerical ranges and parameters setting forth the broad scope of the invention are approximations, The values ​​given in the examples are reported as precisely as possible. However, all values ​​are subject to the There is a certain amount of error necessarily resulting from the standard deviation found in their respective testing measurements. can be.

[0063] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limiting. Each group member may be used either individually or in conjunction with other members of the group. The present invention relates to a method and apparatus for producing a pharmaceutical composition comprising the steps of: For reasons of convenience and / or patentability, one or more members of a group may be grouped together. It is expected that a loop will be included or removed from the group. Any such inclusion or If any such modification or deletion is made, this specification will be deemed to include the modified group and the attached A description of all Markush groups used in the claims is provided.

[0064] Some embodiments of the present invention are the best known to the inventors for carrying out the invention. Of course, variations of these described embodiments may be made without departing from the spirit and scope of the present invention. The forms will be apparent to those of skill in the art upon reading the foregoing description. It is expected that such modifications will be adopted where appropriate, and the inventors specifically refer to them herein as such. It is intended that the invention be practiced in ways not described. To the extent permitted by law, all modifications and equivalents of the subject matter recited in the claims appended hereto. Furthermore, any combination of the above-described elements in all possible variations thereof. Unless otherwise indicated herein or clearly contradicted by context, is included in.

[0065] Numerous patents and publications are referenced throughout this specification. Each of the cited publications is individually incorporated herein by reference in its entirety.

[0066] Moreover, the embodiments of the invention disclosed herein are illustrative of the principles of the invention. It is to be understood that other variations that may be used are within the scope of the present invention. By way of example, and not of limitation, alternative configurations of the present invention may be utilized in accordance with the teachings herein. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

1. A method for culturing a first type of cell, comprising: providing a container containing the first type of cells, the container having an exterior surface and an interior surface, the interior surface comprising a fluoropolymer, the container housing a degradable carrier; the degradable carriers are a plurality of degradable microcarriers having an average diameter in the range of 0.5 μm to 1000 μm, each degradable microcarrier comprising a body formed of a hydrolyzable polymer, the body having a surface and a plurality of cell-trapping moieties attached to the surface of the body; or the degradable carrier is a coating disposed adjacent to the interior surface of the container, the coating including a body formed of a hydrolyzable polymer, the body having a surface and a plurality of cell capture moieties attached to the surface of the body; Each of the cell-trapping moieties is capable of selectively binding to a first type of cell, and the first type of cell is selectively bound to the degradable carrier via the cell-trapping moieties; providing a container, wherein the hydrolyzable polymer is one or more polymers selected from starch, polygalacturonic acid, poly(lactic-co-glycolic acid) (PLGA), polylactic acid (PLA), and polycaprolactone; mixing the cells of the first type with the degradable carrier in a first cell culture medium contained in the container such that the cells of the first type selectively bind to the degradable carrier via the cell-trapping moiety; removing the first cell culture medium from the vessel while retaining the degradable support within the vessel; adding a second cell culture medium to the vessel; and degrading the degradable carrier by hydrolysis of the body of a hydrolyzable polymer, the degradation of the degradable carrier releasing the first type of cells therefrom.

2. The method of claim 1, wherein the degradable carriers are a plurality of degradable microcarriers having an average diameter in the range of 0.5 μm to 1000 μm, each degradable microcarrier comprising a body formed of a hydrolyzable polymer, the body comprising a surface and a plurality of cell-trapping moieties attached to the surface of the body.

3. The method of claim 1, wherein the degradable carrier is a coating disposed adjacent to the inner surface of the container, the coating comprising a body formed of a hydrolyzable polymer, the body having a surface and a plurality of cell capture moieties attached to the surface of the body.

4. The method of claim 1 , wherein each of the cell-capturing moieties comprises one or more aptamers.

5. The method of claim 1 , wherein the cell is a blood cell or an immune cell.

6. The method of claim 5, wherein the cell is a monocyte, a stem or progenitor cell, a T cell, an endothelial progenitor cell, or a natural killer cell.

7. 7. The method of claim 1, wherein the degradable carrier is degraded in water at a pH below 6 or above 8.

8. 7. The method according to any one of claims 1 to 6, wherein the decomposable support is decomposed in water upon contact with a hydrolysis catalyst.

9. 7. The method of claim 1, wherein the degradable carrier is degraded in water upon contact with an enzyme, or an enzyme and a chelating agent.

10. 7. The method of claim 1, wherein the degradable carrier decomposes in water when exposed to a temperature above 40°C.

11. 11. The method according to any one of claims 1 to 10, wherein the fluoropolymer is polytetrafluoroethylene (PTFE), perfluoroalkoxy (PFA), ethylene tetrafluoroethylene (ETFE), polyvinylidene fluoride (PVDF), polychlorotrifluoroethylene (PCTFE), ethylene chlorotrifluoroethylene (ECTFE), fluorinated ethylene propylene (FEP), ethylene fluorinated ethylene propylene (EFEP), perfluoropolyether (PFPE), modified polytetrafluoroethylene (TFM), polyvinyl fluoride (PVF), or any mixture thereof.

12. The method of any one of claims 1 to 11, wherein the provided container has an off-target agent contained therein, the off-target agent being a second plurality of cells of a second type other than the first type, and less than 10% of the off-target agent being bound to the degradable carrier.

13. A container, A rocker configured to mix the degradable carrier with an aqueous medium, a centrifuge configured to separate the degradable carrier from the aqueous medium, and a filter in fluid communication with an outlet of the container and configured to separate the degradable carrier from the aqueous medium; The method according to any one of claims 1 to 12, wherein the method is carried out using a cell isolation system comprising:

14. 14. The method of any one of claims 1, 2, and 4-13, wherein the degradable carriers are a plurality of degradable microcarriers, and removing the first cell culture medium comprises sedimenting the microcarriers in a centrifuge and decanting the first cell culture medium from the container, or removing the first cell culture medium from the container through a filter configured to retain the microcarriers.

15. 15. The method of any one of claims 1 and 3-14, wherein the degradable support is a degradable coating disposed adjacent to the interior surface of the vessel, and removing the first cell culture medium comprises decanting the first cell culture medium from the vessel.

16. the provided container contains a second plurality of cells of a second type; 16. The method of any one of claims 1 to 15, wherein at least 80% of the cells in the vessel after removing the first cell culture medium are the first type of cells.

17. 17. The method according to any one of claims 1 to 16, wherein the step of decomposing the decomposable support comprises a step of subjecting the decomposable support to a pH of less than 6 or more than 8, a step of contacting the decomposable support with a hydrolysis catalyst, a step of contacting the decomposable support with an enzyme and optionally a chelating agent, or a step of exposing the decomposable support to a temperature of more than 40°C.

18. 18. The method of claim 1, wherein degrading the degradable carrier comprises at least partially degrading the degradable carrier.

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