WO2020142453A1 - Containers containing degradable carriers - Google Patents
Containers containing degradable carriers Download PDFInfo
- Publication number
- WO2020142453A1 WO2020142453A1 PCT/US2019/068983 US2019068983W WO2020142453A1 WO 2020142453 A1 WO2020142453 A1 WO 2020142453A1 US 2019068983 W US2019068983 W US 2019068983W WO 2020142453 A1 WO2020142453 A1 WO 2020142453A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- container
- carrier
- biological agent
- degradable
- cell
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/505—Containers for the purpose of retaining a material to be analysed, e.g. test tubes flexible containers not provided for above
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/20—Material Coatings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/30—Constructional details, e.g. recesses, hinges biodegradable
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
- B01L2300/123—Flexible; Elastomeric
Definitions
- This disclosure relates generally to containers (such as bags) containing degradable carriers. More particularly, the present disclosure relates to containers such as bags comprising a fluoropolymer and containing a degradable carrier comprising biological agent-capturing moieties, to cell isolation systems comprising such bags, and to methods for cultivating a biological agent using such bags.
- Cell culture and cell isolation are important processes in a number of applications.
- certain cells for use in therapeutic applications e.g., immunotherapy, regenerative medicine, etc.
- cells such as progenitor cells and mesenchymal stem cells, and monocytes and other immune cells are present in blood in relatively low concentrations, and accordingly are typically isolated from blood and cultured in vitro.
- neuronal cells, cardiomyocytes, epithelial cells, and other cells for regenerative medicine e.g., bone repair, skin repair, pancreatic islets regeneration, etc.
- regenerative medicine e.g., bone repair, skin repair, pancreatic islets regeneration, etc.
- Fluoropolymer bags are commonly used for cell cultures. Such bags are typically inexpensive, disposable, portable and easy to use. Conventionally, cells and feed medium are mixed within the culture bag. As nutrients are consumed and cell waste accumulates, the feed medium must be removed and replenished. Because cells can remain suspended within the bag, the desired cells can be removed along with the depleted medium.
- T cells which play a central role in cell-mediumted immunity
- TCR T cell receptor
- a solid support having attached proteins capable of binding T cell receptors will selectively capture T cells within a cell culture, while undesired cells remain suspended.
- releasing captured cells for example, by cleaving the anchoring protein, can undesirably alter the isolated cells.
- the disclosure provides a container (e.g., in the form of a bag) having an outer surface and an inner surface, the inner surface comprising a fluoropolymer. and contained in the container, a degradable carrier; wherein the carrier comprises a plurality of biological agent-capturing moieties.
- the disclosure provides a method for cultivating a biological agent, comprising providing a container as described herein, the container containing the biological agent; mixing the biological agent and the carrier in a first aqueous medium contained in the container; removing first aqueous medium from the container; adding a second aqueous medium to the container; and degrading the carrier.
- Fig. 1 is a schematic top-down (top) and cross-sectional (bottom) view of a bag according to one embodiment of the disclosure.
- Fig. 2 is a schematic top-down view of bags according to certain embodiments of the disclosure.
- Fig. 3 is a schematic cross-sectional view of a bag according to one embodiment of the disclosure.
- Fig. 4 is a schematic cross-sectional view of a bag according to one embodiment of the disclosure.
- the disclosure relates to containers (e.g., bags) having an inner surface comprising a fluoropolymer and containing a degradable carrier comprising biological agentcapturing moieties.
- containers e.g., bags
- a degradable carrier comprising biological agentcapturing moieties.
- desired cells can be captured and physically separated from a suspension within the container, and can further be
- the containers of the disclosure can be provided in a number of forms.
- One especially convenient form is a bag, e.g., formed from one or more sheets of fluoropolymeric material as described herein.
- the person of ordinary skill in the art will be familiar with bag structures such as those used in cell culture, and will be able to adapt conventional bag structures for use in bags and methods of the disclosure based on the description herein
- the person of ordinary skill in the art will appreciate that the containers of the disclosure can be provided in a number of other forms, e.g., flasks, tubes, dishes.
- one aspect of the disclosure is a container, e.g., in the form of a bag having an outer surface and an inner surface, the inner surface comprising a fluoropolymer.
- An embodiment of such a bag is shown in schematic top-down view (top) and cross- sectional view (bottom) in Fig. 1.
- Bag 100 of Fig. 1 includes a bag wall 110 having an outer surface 112 and an inner surface 114, and further includes ports 130 and 140, located at opposite ends of the bag for adding or removing medium (e.g., feed medium or waste medium, respectively) to or from the bag.
- medium e.g., feed medium or waste medium, respectively
- the person of ordinary skill in the art will appreciate that the number and location of ports are not particularly limited, and accordingly can be positioned, for example, for convenience of use or manufacture.
- Bag 100 can be the product of bonding two fluoropolymer-containing sheets (e.g., two sheets having a layer of fluorinated ethylene propylene sheets on an inside surface thereof) together at their edges (e.g., by laser welding, corona discharge, radiation, heat or melt lamination, etching, plasma treatment, wetting, adhesives, or combinations thereof) to form compartment 120.
- Ports 130 and 140 can be sealable to provide a sealed compartment 120.
- the container wall can be uniform in its composition, or alternatively can include two or more distinct domains (e.g., two or more layers).
- bonding two fluoropolymer sheets together, then coating the bonded sheets can provide an outer surface 112 differing in composition from inner surface 114.
- bonding two multi-layer sheets together can provide an outer surface 112 differing in composition from inner surface 114.
- Multilayer sheets can be formed of both fluorpolymeric and nonfluorinated polymer materials; in such cases, a fluoropolymer layer can be provided at the inner surfaces of one or more of the multi-layer sheets.
- the thickness of the container, the volume of the compartment, and the shape of the container and/or compartment are not particularly limited, and can be selected for convenience of use or manufacture, and/or to suit a specific application.
- the thickness of the container wall can be within the range of 0.0003 inches to 0.2 inches
- the volume of the compartment can be within the range of 100 mL to 100 L.
- FIG. 2 shows several exemplary embodiments of configurations for culture bags suitable for use in the bags and methods of the disclosure.
- Bag 200a has only a single port 230a, providing access to compartment 220a.
- Bag 200b is in the so-called“serpentine" configuration, in which a longer path length through the system can be provided; ports 230b and 240b are connected by a serpentine path formed by serpentine-shaped compartment 220b formed by appropriate welding of the sheets forming the bag.
- bag 200c has a non-rectangular shape, with a corresponding non-rectangular compartment 220c between ports 230c and 240c.
- One or more of the walls of the container can be porous, and can, for example, be permeable to gases produced and consumed in a cell culture (e.g., O 2 , CO 2 ) but impermeable to liquids (e.g., water). This can allow for passive exchange of gases across the container walls with the atmosphere to allow for respiration of cells in the bag.
- a cell culture e.g., O 2 , CO 2
- liquids e.g., water
- the containers of the disclosure are desirably formed such that there is substantially no contamination of a fluid within the container. Accordingly, it is desirable for the inner surface of the container to be formed from materials that will not leach organics into the fluid.
- an inner surface of the container wall is formed of a polymer (e.g., a fluoropolymer such as fluorinated ethylene propylene) having a total organic carbon (TOC) in water of less than 0.1 mg/cm 2 (e.g., less than 0.05 mg/cm 2 , or less than 0.05 mg/cm 2 ).
- a polymer e.g., a fluoropolymer such as fluorinated ethylene propylene
- TOC total organic carbon
- containers of the disclosure need not have reactive functional groups at the inner surfaces thereof, although in some cases, such as when the degradable carrier is a degradable coating, reactive functional groups can be used to covalently attach the degradable carrier to the inner surface of the container as described below.
- the container contains a degradable earner, and the carrier comprises a plurality of biological agent-capturing moieties.
- a biological agent-capturing moiety is a material with which the biological agent can selectively associate. The association between a biological agent and a respective capturing moiety is sufficient to retain the biological agent on the carrier (e.g., throughout centrifugation or filtration), and may be covalent or non-covalent.
- the biological agent-capturing moiety comprises one or more aptamers.
- the one or more aptamers are selected from peptides, polypeptides, oligonucleotides, and polynucleotides (i.e., DNA and RNA).
- the biological agent-capturing moiety comprises an oligonucleotide aptamer selected from a random, synthetically generated sequence pool.
- the biological agentcapturing moiety comprises a DNA aptamer having at least two sites to which the biological agent can selectively associate. For example, in certain such embodiments, at least 5, or at least 10, or at least 25, or at least 50, or at least 75, or at least 100 instances of a nucleotide subsequence selective for the biological agent are present in the DNA aptamer sequence.
- the biological agent- capturing moiety comprises a plurality of aptamers (e.g., oligonucleotide aptamers) attached to a polypeptide.
- the biological agent-capturing moiety comprises a plurality of biotinylated aptamers attached to an avidin protein.
- the biotinylated aptamers comprise oligonucleotides or polypeptides.
- the biological agentcapturing moiety comprises at least 10, or at least 25, or at least 50, or at least 75, or 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 700, or at least 800, or at least 900, or at least 1000 aptamers (e.g., biotinylated aptamers attached to an avidin protein).
- aptamers e.g., biotinylated aptamers attached to an avidin protein.
- the biological agentcapturing moiety comprises a number of aptamers (e.g., biotinylated aptamers attached to an avidin protein) within the range of 10 to 1000, or 10 to 900, or 10 to 800, or 10 to 700, or 10 to 600, or 10 to 500, or 10 to 400, or 10 to 300, or 10 to 200, or 25 to 1000, or 50 to 1000, or 100 to 1000, or 200 to 1000, or 300 to 1000, or 400 to 1000, or 500 to 1000, or 600 to 1000, or 700 to 1000, or 800 to 1000, or 10 to 400, or 100 to 500, or 200 to 600, or 300 to 700, or 400 to 800, or 500 to 900, or 600 to 1000.
- aptamers e.g., biotinylated aptamers attached to an avidin protein
- the biological agent is selected from inorganic species and organic small molecules.
- the biological agent is a metal ion.
- the biological agent is a vitamin, hormone, or peptide.
- the biological agent is a macromolecule such as, for example, a protein, an enzyme, or a nucleic acid (i.e., DNA or RNA).
- the biological agent is complex biological system such as, for example, a cell organelle (e.g., nucleus, ribosome, mitochondria, vacuole, rough endoplasmic reticulum, smooth endoplasmic reticulum, Golgi apparatus, lysosome, centrosome, vesicle, membrane) or a cell fragment.
- a cell organelle e.g., nucleus, ribosome, mitochondria, vacuole, rough endoplasmic reticulum, smooth endoplasmic reticulum, Golgi apparatus, lysosome, centrosome, vesicle, membrane
- the biological agent is a bacterium or a cell group.
- the biological agent is a cell (e.g., a normal cell or a diseased cell).
- the cell is a blood cell, a stem cell, or an immune cell.
- the cell is a white blood cell.
- the cell is a monocyte.
- the cell is a stem cell such as, for example, a mesenchymal stem cell, or a progenitor cell.
- the cell is a T cell (e.g., a regulatory T cell), an endothelial progenitor cell, or a natural killer cell.
- the biological agent can, in certain embodiments, be a desirable biological agent, capturable for retention in a cell culture (i.e., positive selection), and can alternatively, in other embodiments, be an undesirable biological agent, capturable for removal from a cell culture (i.e., negative selection).
- the biological agent-capturing moiety comprises one or more aptamers that can selectively associate with the CD31 marker of an endothelial progenitor cell.
- the biological agent-capturing moiety comprises one or more aptamers that can selective associate with one or more markers of a T regulatory cell (e.g., CD4, CD25, CD27).
- the biological agent-capturing moiety comprises one or more aptamers that can selectively associate with one or more markers of a mesenchymal stem cell (e.g., CD73, CD90, CD105).
- the biological agentcapturing moiety comprises a plurality of aptamers (e.g., oligonucleotide aptamers) attached to a polypeptide.
- a degradable carrier is a material having in an undegraded state a surface to which the biological agent-capturing moieties can be attached (e.g., through a covalent linker).
- the carrier can remain in an undegraded state in the presence of, for example, an aqueous medium for at least a period of time sufficient to culture or isolate a biological agent, or even indefinitely.
- the carrier can be degraded into fragments or can be partially or completely dissolved, and in a degraded state can be separated from the biological agent.
- the biological agentcapturing moiety is covalently linked to a functional group of the surface of the degradable carrier.
- the surface of the degradable carrier may be functionalized with a carboxyl group, hydroxyl group, aldehyde group, carbonyl group, amine group, imine group, amide group, ester group, anhydride group, thiol group, disulfide, phenol, guanidine, thioether, indole, imidazole, or diazonium group.
- the biological agent-capturing moiety comprises a plurality of aptamers (e.g., oligonucleotide aptamers) attached to a protein, the protein attached through a peptide linkage to a carboxyl group of the surface of the degradable carrier.
- the biological agent-capturing moiety comprises an amine-terminated aptamer (e.g., a DNA aptamer) attached through an imine linkage to an aldehyde group of the surface of the degradable carrier.
- the number of biological agent-capturing moieties per unit of degradable carrier surface is not particularly limited, the person of ordinary skill in the art will appreciate that the capturing moiety density can, typically, be greater than the number of biological agents (e.g., cells) that can be captured per unit of degradable carrier surface (i.e., captured agent density). Accordingly, in certain embodiments, the capturing moiety density of the degradable carrier is greater than the captured agent density of the degradable carrier.
- the degradable carrier comprises a plurality of degradable microcarriers (e.g., polymer microcarriers), the microcarriers having a surface to which the biological agent-capturing moieties can be attached.
- bag 300 of Fig. 3 includes a bag wall 310 (having an outer surface 312 and an inner surface 314) enclosing a compartment 320, compartment 320 containing a plurality of degradable microcarriers 350.
- the average size of the microcarriers is within the range of 0.5 mm to 1000 mm, or 0.5 mm to 900 mm, or 0.5 mm to 800 mm, or 0.5 mm to 700 mm, or 0.5 mm to 600 mm, or 0.5 mm to 500 mm, or 0.5 mm to 400 mm, or 0.5 mm to 300 mm, or 0.5 mm to 250 mm, or 1 mm to 1000 mm, or 5 mm to 1000 mm, or 10 mm to 1000 mm, or 25 mm to 1000 mm, or 50 mm to 1000 mm, or 100 mm to 1000 mm, or 200 mm to 1000 mm, or 300 mm to 1000 mm, or 400 mm to 1000 mm, or 500 mm to 1000 mm, or 10 mm to 750 mm, or 10 mm to 500 mm, or 20 mm to 400 mm, or 50 mm to 300 mm.
- the degradable carrier comprises a degradable coating (e.g., a polymer coating) disposed adjacent to at least a portion (e.g., at least a majority, or even all) of the inner surface of the container.
- the degradable coating is covalently attached to the inner surface of the container.
- bag 400 of Fig. 4 includes a bag wall 410 (having an outer surface 412 and an inner surface 414) enclosing a sealed compartment 420, sealed compartment 420 containing a degradable coating 460 disposed adjacent to (e.g., covalently attached to) inner surface 414.
- the degradable coating has a thickness within the range of 10 nm (e.g., about a monolayer) to 10 mm, or 10 nm to 1 mm, or 10 nm to 500 mm, or 10 nm to 100 mm, or 10 nm 1 mm, or 1 mm to 10 mm, or 100 mm to 10 mm, or 500 mm to 10 mm, or 1 mm to 10 mm.
- the degradable coating is covalently linked to a functional group of the inner surface of the container (e.g., a functional group of the fluoropolymer comprising the inner surface).
- the functional group is a carboxyl group, hydroxyl group, aldehyde group, carbonyl group, amine group, imine group, amide group, ester group, anhydride group, thiol group, disulfide, phenol, guanidine, thioether, indole, imidazole, or diazonium group.
- the degradable carrier comprises a cleavable polymer.
- the degradable carrier comprises an enzyme-cleavable polymer such as, for example, a polysaccharide.
- the polysaccharide is a starch (e.g., deavable by alpha- amylase).
- the polysaccharide is a polygalacturonic acid (e.g., deavable by pedinase).
- the degradable carrier comprises a hydrolyzable polymer such as, for example, poly(lact -co-glycolic acid) (PLGA), polyladic acid (PLA), or polycaproladone.
- PLGA poly(lact -co-glycolic acid)
- PLA polyladic acid
- polycaproladone polycaproladone
- the polymer hydrolysis is accelerated at a pH below 6, or at a pH above 8.
- the polymer hydrolysis is accelerated in the presence of a hydrolysis catalyst such as, for example, an amine.
- the degradable carrier comprises a cross-linked polymer having, for example, a number of cross-links suffident to provide a carrier that can remain undegraded in an aqueous medium.
- the degradable carrier comprises an enzyme-deavable polymer (e.g., a polysaccharide) having a number of cross-links (e.g., glycerol diether cross-links) suffident to provide a carrier that can remain undegraded in an aqueous medium having less than a critical level of the enzyme.
- the degradable carrier comprises a hydrolyzable polymer (e.g., PLGA, PLA, or polycaproladone) having a number of cross-links suffident to provide a carrier that can remain undegraded in an aqueous medium having less than a critical level of a hydrolysis catalyst, or in an aqueous medium having a pH outside of a critical range.
- the degradable carrier comprises an insoluble cross-linked network of an otherwise soluble polymer.
- a degradable carrier comprising a thioether-cross-linked polymer network (e.g., a hydrogel) can remain undegraded in an aqueous medium having less than a critical level of glutathione.
- a thioether-cross-linked polymer network e.g., a hydrogel
- a cross-linked polymer of the degradable carrier comprises a cross-linker that prevents enzyme cleavage of the polymer (e.g., of a polysaccharide).
- a degradable carrier comprising a Ca 2+ -cross-linked polysaccharide e.g., polygalacturonic acid
- a degradable carrier comprising a Ca 2+ -cross-linked polysaccharide can remain undegraded in an aqueous medium having less than a critical level of a Ca 2+ -chelating agent (e.g., EDTA), i.e., even if the medium includes more than a critical level of a cleaving enzyme (e.g., pectinase).
- a critical level of a cleaving enzyme e.g., pectinase
- the degradable carrier is degradable upon, or the degradation of the degradable carrier is accelerated upon, irradiation (e.g., with radiation having a wavelength of less than 450 nm, or less than 420 nm).
- the degradable carrier is degradable upon, or the degradation of the degradable carrier is accelerated upon, exposure to an elevated temperature (e.g., greater than 37 °C, or greater than 40 °C).
- the inner surface of the container comprises a fluoropolymer selected from polytetrafluoroethylene (PTFE), perfluoroalkoxy (PFA), ethylene tetrafluoroethylene (ETFE), polyvinylidene fluoride (PVDF), polychlorotrifluoroethylene (PCTFE), ethylene chlorotrifluoroethylene (ECTFE), fluorinated ethylene propylene (FEP), ethylene fluorinated ethylene propylene (EFEP),
- PTFE polytetrafluoroethylene
- PFA perfluoroalkoxy
- ETFE ethylene tetrafluoroethylene
- PVDF polyvinylidene fluoride
- PCTFE polychlorotrifluoroethylene
- ECTFE ethylene chlorotrifluoroethylene
- FEP ethylene fluorinated ethylene propylene
- the inner surface of the container comprises fluorinated ethylene propylene.
- the inner surface of the container consists essentially of, or is the fluoropolymer (e.g., fluorinated ethylene propylene).
- the material comprising the inner 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, at least 0.001 inches, or at least 0.10 inches.
- the material at the inner surface of the container has a thickness within the range of 0.0003 inches to 0.2 inches, or 0.0003 inches to 0.1 inches, or 0.0005 inches to 0.08 inches, or 0.001 inches to 0.07 inches, or 0.001 inches to 0.05 inches, or 0.001 inches to 0.03 inches, or 0.001 inches to 0.018 inches, or 0.001 inches to 0.016 inches, or 0.001 inches to 0.014 inches, or 0.001 inches to 0.012 inches.
- the material making up the container wall is a multilayer material, with a layer of fluoropolymer at the inner surface thereof, and a layer of another polymeric material (fluoro polymeric or otherwise) at the outer surface thereof.
- the material at the outer surface of the container has a thickness of at least 0.0005 inches, or at least 0.001 inches, or at least 0.005 inches, or at least 0.0075 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 0.05 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 inches, or at least 0.11 inches.
- the material at the outer surface of the container has a thickness within the range of 0.0005 inches to 0.2 inches, or 0.005 inches to 0.18 inches, or 0.01 inches to 0.16 inches, or 0.01 inches to 0.14 inches, or 0.01 inches to 0.12 inches, or 0.06 inches to 0.13 inches, or 0.09 inches to 0.126 inches.
- the degradable carrier comprises a plurality of microcarriers and the inner surface of the container further comprises a plurality of biological agent-capturing moieties as described herein.
- the biological agent-capturing moieties attached to the inner surface of the container are the same as the biological agent-capturing moieties of the degradable carrier.
- the biological agent-capturing moieties attached to the inner surface of the container are selective for a biological agent other than the agent for which the biological agent-capturing moieties of the degradable carrier are selective.
- the present inventors have determined that such containers can improve biological agent capture efficiency, or can facilitate the isolation and separation of two distinct biological agents.
- the outer surface of the container comprises a material other than a fluoropolymer.
- the material of the outer surface of the container comprises a thermoplastic polymer, a thermoplastic elastomer, a silicon, a rubber, or any combination thereof.
- the outer surface of the container can, in certain embodiments as otherwise described herein, comprise a fluoropolymer such as, for example, the fluoropolymer of the inner surface.
- the material of the inner surface and the outer surface i.e., the container wall
- the fluoropolymer e.g., fluorinated ethylene propylene
- the container includes the biological agent.
- the biological agent is attached to the carrier through the biological agent-capturing moiety.
- the biological agent is a cell associated with the carrier through an aptamer bound to the surface of a degradable microcarrier, or a degradable coating.
- the inner surface of the container comprises a plurality of the biological agent-capturing moieties, at least a portion of which are attached to the biological agent.
- the biological agent is a cell, and can associate with an aptamer bound to the surface of a degradable microcarrier, or with an aptamer bound to the inner surface of the container.
- the biological agent-capturing moiety of the degradable microcarriers is the same as the biological agent-capturing moiety at the inner surface of the container.
- the container further includes an aqueous medium.
- the aqueous medium can be, for example, a cell culture medium comprising a biological agent and one or more off-target agents.
- off-target agents are materials other than the biological agent (i.e., capturable by the biological agent-capturing moiety) including, for example, cells other than the biological agent, cell fragments, proteins, vitamins, hormones, peptides, and metal ions.
- the container includes an off-target agent (e.g., a cell other than the biological agent), and no more than 20% of the off-target agent is adhered to the degradable carrier or the inner surface of the container.
- an off-target agent e.g., a cell other than the biological agent
- the container includes an off-target agent (e.g., a cell other than the biological agent), and no more than 20% of the off-target agent is adhered to the degradable carrier or the inner surface of the container.
- the off-target agent e.g., a cell other than the biological agent
- the cell isolation system includes a container as otherwise described herein.
- the cell isolation system comprises a container as otherwise described herein containing an aqueous medium, and a rocker configured to mix the aqueous medium and the carrier.
- the cell isolation system comprises a container as otherwise described herein containing an aqueous medium, and a centrifuge configured to separate the carrier (e.g., comprising a plurality of degradable microcarriers) from the aqueous medium.
- the cell isolation system comprises a container as otherwise described herein containing an aqueous medium, and a filter in fluid communication with an outlet of the container configured to separate the carrier (e.g., comprising a plurality of degradable microcarriers) from the aqueous medium.
- the aqueous medium in the container of the cell isolation system includes an off-target agent such as, for example, a cell other than the biological agent.
- the present inventors have determined that degradation of the carrier described herein can release a captured (e.g., and isolated) biological agent into aqueous medium (e.g., to be stored or further cultured) without deleteriously affecting cell function.
- a captured biological agent e.g., and isolated
- the fluoropolymer of the inner surface can prevent adhesion of materials other than the biological agent (i.e., off-target agents), increasing the purity of a biological agent isolated in the container.
- another aspect of the disclosure is a method for cultivating a biological agent, the method including providing a container as otherwise described herein containing the biological agent, and mixing the biological agent and the carrier in a first aqueous medium contained in the container. The method further includes removing the first aqueous medium from the container, adding a second aqueous medium to the container, and degrading the carrier.
- the container contains a blood sample (e.g., a whole blood sample) including the biological agent.
- the container contains a blood sample and the biological agent is a monocyte.
- the container further includes one or more off-target agents.
- the container contains a sample (e.g., a whole blood sample) including the biological agent (e.g., a monocyte, a stem cell, a T cell, an endothelial progenitor cell, or a natural killer cell) and one or more off-target agents (e.g., a cell other than the biological agent).
- the first aqueous medium is a cell culture medium.
- the biological agent is a cell, and the biological agent is cultured in the container (i.e., the agent is grown, or expanded, in the first aqueous medium).
- one or more off-target agents comprising the aqueous medium may also expand in the first aqueous medium.
- the biological agent and carrier are mixed on a rocker. The mixing is performed for a period of time sufficient to allow the biological agent-capturing moiety and the biological agent to associate.
- the biological agent is a cell, and the mixing is performed for a period of time and at a temperature sufficient to expand the biological agent. In certain embodiments, the mixing is performed on a rocker.
- the carrier comprises a plurality of degradable microcarriers
- removing the first aqueous medium comprises sedimenting the carriers and decanting the first aqueous medium from the container.
- removing the first aqueous medium comprises sedimenting the microcarriers with a centrifuge and decanting the first aqueous medium from the container.
- the first aqueous medium is removed from the container through a filter configured to retain the microcarriers.
- the first aqueous medium is removed from the container through a filter having a pore size smaller than the degradable microcarriers in an undegraded state.
- the carrier comprises a degradable coating disposed adjacent the inner surface of the container, and removing the first aqueous medium comprises decanting the first aqueous medium from the container (i.e., without
- the container as otherwise described herein can contain cells including, for example, the biological agent and optionally one or more off-target agents.
- a majority of the cells remaining in the container can comprise the biological agent attached to the degradable carrier and optionally to the inner surface of the container through the biological agent-capturing moiety.
- the biological agent is a cell, and at least 80%, 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% of the cells in the container after removing the first aqueous medium are the biological agent.
- degrading the carrier comprises subjecting the carrier to a pH different than the pH of the first aqueous medium.
- degrading the carrier comprises subjecting the carrier to a pH below 6 or above 8.
- the second aqueous medium added to the container has a pH below 6 or above 8.
- the pH of the second aqueous medium is adjusted after adding the second aqueous medium to the container.
- degrading the carrier comprises contacting the carrier with a hydrolysis catalyst (e.g., an amine catalyst).
- the second aqueous medium added to the container contains a hydrolysis catalyst.
- a hydrolysis catalyst is added to the container after adding the second aqueous medium.
- the pH of the second aqueous medium enables, or even accelerates the hydrolysis reaction catalyzed by the hydrolysis catalyst.
- degrading the carrier comprises contacting the carrier with an enzyme, or an enzyme and a chelating agent.
- the second aqueous medium added to the container contains an enzyme.
- an enzyme is added to the container after adding the second aqueous medium.
- degrading the carrier comprises subjecting the carrier to an elevated temperature (e.g., a temperature greater than 37 °C, or greater than 40 °C).
- degrading the carrier comprises irradiating the carrier (e.g., with radiation having a wavelength of less than 450 nm, or less than 420 nm).
- the present inventors have determined that degrading (e.g., fragmenting, partially dissolving, or dissolving) the carrier can separate a captured biological agent from the carrier without deleteriously affecting the biological agent.
- the method further comprises culturing the biological agent in the second aqueous medium (i.e., including the degraded carrier).
- the second aqueous medium is a cell culture medium.
- the method further comprises removing the degraded carrier, for example by sedimenting the biological agent and removing the second aqueous medium, and adding a third aqueous medium to the container.
- the third aqueous medium is a cell culture medium, or a cell storage medium.
- Embodiment 1 A container (e.g., in the form of a bag) having an outer surface and an inner surface, the inner surface comprising a fluoropolymer, and contained in the container, a degradable carrier;
- a container e.g., in the form of a bag having an outer surface and an inner surface, the inner surface comprising a fluoropolymer, and contained in the container, a degradable carrier;
- the carrier comprises a plurality of biological agent-capturing moieties.
- Embodiment 2 The container of embodiment 1 , wherein the degradable carrier comprises a plurality of degradable microcarriers, each microcarrier comprising a plurality of biological agent-capturing moieties.
- Embodiment 3 The container of embodiment 2, wherein the average diameter of the microcarriers is within the range of about 0.5 mm to about 1000 mm (e.g., about 5 mm to about 750 mm, or about 10 mm to about 500 mm, or about 25 mm to about 300 mm).
- Embodiment 4 The container of embodiments 2 or 3, wherein the inner surface of the container comprises a plurality of biological agent-capturing moieties.
- Embodiment 5 The container of embodiment 1 , wherein the degradable carrier comprises a degradable coating disposed adjacent to at least a portion (e.g., at least a majority) of the inner surface of the container.
- Embodiment 6 The container of any of embodiments 1-5, wherein the biological agent-capturing moiety comprises one or more aptamers.
- Embodiment 7. The container of any of embodiments 1-6, wherein the biological agent is a cell.
- Embodiment 8 The container of embodiment 7, wherein the cell is a blood cell or an immune cell.
- Embodiment 9 The container of embodiment 7, wherein the cell is a monocyte, a stem or progenitor cell (e.g., a mesenchymal stem cell), a T cell (e.g., a regulatory T cell), an endothelial progenitor cell, or a natural killer cell.
- a monocyte e.g., a monocyte
- a stem or progenitor cell e.g., a mesenchymal stem cell
- a T cell e.g., a regulatory T cell
- an endothelial progenitor cell e.g., a natural killer cell.
- Embodiment 10 The container of any of embodiments 1-9, wherein the degradable carrier comprises a polymer.
- Embodiment 11 The container of embodiment 10, wherein the degradable carrier comprises a polysaccharide (e.g., starch or polygalacturonic acid).
- a polysaccharide e.g., starch or polygalacturonic acid
- Embodiment 12 The container of embodiment 10, wherein the degradable carrier comprises one or more polymers selected from poly(lactic-co-glycolic acid) (PLGA), polylactic acid (PLA), and polycaprolactone.
- PLGA poly(lactic-co-glycolic acid)
- PLA polylactic acid
- PCAP polycaprolactone
- Embodiment 13 The container of any of embodiments 10-12, wherein the polymer is crosslinked (e.g., thioether cross-linked, Ca 2+ cross-linked, or glycerol diether cross-linked).
- crosslinked e.g., thioether cross-linked, Ca 2+ cross-linked, or glycerol diether cross-linked.
- Embodiment 14 The container of any of embodiments 1-13, wherein the carrier is degradable in water at a pH below 6, or at a pH above 8.
- Embodiment 15 The container of any of embodiments 1-13, wherein the carrier is degradable in water upon contact with a hydrolysis catalyst (e.g., an amine catalyst).
- a hydrolysis catalyst e.g., an amine catalyst
- Embodiment 16 The container of any of embodiments 1-13, wherein the carrier is degradable in water upon contact with an enzyme, or an enzyme and a chelating agent.
- Embodiment 17 The container of embodiment 16, wherein the enzyme is an amylase or a pectinase.
- Embodiment 18 The container of any of embodiments 1-13, wherein the carrier is degradable in water upon exposure to an elevated temperature (e.g., a temperature greater than 37 °C, or greater than 40 °C).
- Embodiment 19 The container of any of embodiments 1-13, wherein the carrier is degradable in water upon irradiation (e.g., with radiation having a wavelength of less than 450 nm, or less than 420 nm).
- Embodiment 20 The container of any of embodiments 1-19, 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.
- PTFE polytetrafluoroethylene
- PFA perfluoroalkoxy
- ETFE ethylene tetrafluoroethylene
- PVDF polyvinylidene fluoride
- PCTFE polychlorotrifluoroethylene
- ECTFE ethylene chlorotrifluoroethylene
- FEP
- Embodiment 21 The container of any of embodiments 1-19, wherein the fluoropolymer is fluorinated ethylene propylene (FEP).
- FEP fluorinated ethylene propylene
- Embodiment 22 The container of any of embodiments 1-21 , wherein the degradable carrier is attached to the biological agent, the biological agent attached to the carrier through the biological agent-capturing moiety.
- Embodiment 23 The container of any of embodiments 1-22, containing an aqueous medium.
- Embodiment 24 The container of embodiment 23, containing an off-target agent.
- Embodiment 25 The container of embodiment 24, wherein the off-target agent is a cell other than the biological agent.
- Embodiment 26 The container of embodiment 24 or 25, wherein less than 20% (e.g., less than 15%, less than 10%, less than 5%, or less than 1%) of the off-target agent is adhered to the carrier or the inner surface of the container.
- Embodiment 27 A cell isolation system comprising the container of any of
- Embodiment 28 A cell isolation system comprising the container of any of embodiments 23-25 and a centrifuge configured to separate the carrier from the aqueous medium.
- Embodiment 29 A cell isolation system comprising the container of any of embodiments 23-25 and a filter in fluid communication with an outlet of the container, the filter configured to separate the carrier from the aqueous medium.
- Embodiment 30 A method for cultivating a biological agent, comprising
- Embodiment 31 The method of embodiment 30, wherein the carrier comprises a plurality of degradable microcarriers, and wherein removing the first aqueous medium comprises sedimenting the microcarriers with a centrifuge and decanting the first aqueous medium from the container.
- Embodiment 32 The method of embodiment 30, wherein the carrier comprises a plurality of degradable microcarriers, and wherein the first aqueous medium is removed from the container through a filter configured to retain the microcarriers.
- Embodiment 33 The method of embodiment 30, wherein the carrier comprises a degradable coating disposed adjacent the inner surface of the container, and wherein removing the first aqueous medium comprises decanting the first aqueous medium from the container.
- Embodiment 34 The method of any of embodiments 30-34, wherein
- the biological agent is a cell
- Embodiment 35 The method of any of embodiments 30-34, wherein degrading the carrier comprises subjecting the carrier to a pH below 6 or about 8.
- Embodiment 36 The method of any of embodiments 30-34, wherein degrading the carrier comprises contacting the carrier with a hydrolysis catalyst (e.g., an amine catalyst).
- a hydrolysis catalyst e.g., an amine catalyst
- Embodiment 37 The method of any of embodiments 30-34, wherein degrading the carrier comprises contacting the carrier with an enzyme and, optionally, a chelating agent.
- Embodiment 38 The method of any of embodiments 30-34, wherein degrading the carrier comprises exposing the carriers to an elevated temperature (e.g., a temperature greater than 37 °C, or greater than 40 °C).
- an elevated temperature e.g., a temperature greater than 37 °C, or greater than 40 °C.
- Embodiment 39 The method of any of embodiments 30-34, wherein degrading the carrier comprises irradiating the carriers (e.g., with radiation having a wavelength of less than 450 nm, or less than 420 nm).
- Embodiment 40 The method of any of embodiments 30-39, further comprising removing the degraded carriers from the container.
- Embodiment 41 The container or method of any of claims 1-40, wherein the container is in the form of a bag.
- each embodiment disclosed herein can comprise, consist essentially of or consist of its particular stated element, step, ingredient or component.
- the transition term "comprise” or “comprises” means includes, but is not limited to, and allows for the inclusion of unspecified elements, steps, ingredients, or components, even in major amounts.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Sustainable Development (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Clinical Laboratory Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP19907890.8A EP3906294A4 (en) | 2018-12-31 | 2019-12-30 | Containers containing degradable carriers |
JP2021538428A JP2022517554A (en) | 2018-12-31 | 2019-12-30 | Container containing degradable carrier |
KR1020217024220A KR102615834B1 (en) | 2018-12-31 | 2019-12-30 | Container for receiving degradable carrier |
CN201980087431.7A CN113227344A (en) | 2018-12-31 | 2019-12-30 | Container containing degradable carrier |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862786913P | 2018-12-31 | 2018-12-31 | |
US62/786,913 | 2018-12-31 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2020142453A1 true WO2020142453A1 (en) | 2020-07-09 |
Family
ID=71122728
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2019/068983 WO2020142453A1 (en) | 2018-12-31 | 2019-12-30 | Containers containing degradable carriers |
Country Status (6)
Country | Link |
---|---|
US (1) | US20200209233A1 (en) |
EP (1) | EP3906294A4 (en) |
JP (2) | JP2022517554A (en) |
KR (1) | KR102615834B1 (en) |
CN (1) | CN113227344A (en) |
WO (1) | WO2020142453A1 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113227344A (en) * | 2018-12-31 | 2021-08-06 | 美国圣戈班性能塑料公司 | Container containing degradable carrier |
JP2023511024A (en) * | 2019-12-30 | 2023-03-16 | サン-ゴバン パフォーマンス プラスティックス コーポレイション | Containers and methods for cell transduction |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050196828A1 (en) * | 2002-08-27 | 2005-09-08 | Zheng Ming H. | Bioreactor with expandable surface area for culturing cells |
US20060134790A1 (en) * | 2003-01-13 | 2006-06-22 | Yasunobu Tanaka | Solid surface with immobilized degradable cationic polymer for transfecting eukaryotic cells |
US20110027889A1 (en) * | 2009-07-28 | 2011-02-03 | Mccarthy Kevin Robert | Synthetic Microcarriers for Culturing Cells |
US20160178491A1 (en) * | 2014-12-22 | 2016-06-23 | Saint-Gobain Performance Plastics Corporation | Capture system of cells and methods |
WO2017169259A1 (en) * | 2016-03-28 | 2017-10-05 | ソニー株式会社 | Cell culture container, cell culture system, cell culture kit and cell culture method |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2003289394A1 (en) * | 2002-12-25 | 2004-07-22 | National Institute Of Advanced Industrial Science And Technology | Cell separation and collection apparatus and separation and collection method |
CN1813059A (en) * | 2003-05-02 | 2006-08-02 | 西格玛-奥尔德利希公司 | Solid phase cell lysis and capture platform |
US20050054101A1 (en) * | 2003-07-17 | 2005-03-10 | Global Cell Solutions Llc | Automated cell culture system and process |
JP2008538929A (en) * | 2005-04-29 | 2008-11-13 | イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニー | Multi-step process for producing therapeutic proteins |
JP5280432B2 (en) * | 2007-04-27 | 2013-09-04 | ヒュンジン ヤン | Cell culture support having increased specific gravity and method for producing the same |
ES2647538T3 (en) * | 2007-09-28 | 2017-12-22 | Pfizer Inc. | Addressing to cancer cells using nanoparticles |
JP5309305B2 (en) * | 2008-01-07 | 2013-10-09 | 福岡県 | Culture bag and cell culture method |
US9080933B2 (en) * | 2009-11-09 | 2015-07-14 | University Of Washington Through Its Center For Commercialization | Stimuli-responsive polymer diagnostic assay comprising magnetic nanoparticles and capture conjugates |
WO2014127314A1 (en) * | 2013-02-15 | 2014-08-21 | Phynexus, Inc. | Devices and methods for purification of biological cells |
WO2012158108A1 (en) * | 2011-05-16 | 2012-11-22 | Ge Healthcare Bio-Sciences Ab | Method of cultivating cells on microcarriers in a bag |
WO2015010019A1 (en) * | 2013-07-18 | 2015-01-22 | The General Hospital Corporation | Selective capture and release of rare mammalian cells using photodegradable hydrogels in a microfluidic platform |
US10655097B2 (en) * | 2014-12-22 | 2020-05-19 | Saint-Gobain Performance Plastics Corporation | T-cell culture double bag assembly |
US20160178490A1 (en) * | 2014-12-22 | 2016-06-23 | Saint-Gobain Performance Plastics Corporation | Capture system of cells and methods |
KR101985914B1 (en) * | 2014-12-25 | 2019-06-04 | 도요세이칸 그룹 홀딩스 가부시키가이샤 | Culture container and method for manufacturing culture container |
AU2016338907B2 (en) * | 2015-10-13 | 2022-07-07 | President And Fellows Of Harvard College | Systems and methods for making and using gel microspheres |
US20200108391A1 (en) * | 2017-04-14 | 2020-04-09 | University Of Twente | Device and Method For The Continuous Trapping of Circulating Tumor Cells |
CN113227344A (en) * | 2018-12-31 | 2021-08-06 | 美国圣戈班性能塑料公司 | Container containing degradable carrier |
-
2019
- 2019-12-30 CN CN201980087431.7A patent/CN113227344A/en active Pending
- 2019-12-30 US US16/730,660 patent/US20200209233A1/en active Pending
- 2019-12-30 EP EP19907890.8A patent/EP3906294A4/en active Pending
- 2019-12-30 WO PCT/US2019/068983 patent/WO2020142453A1/en unknown
- 2019-12-30 JP JP2021538428A patent/JP2022517554A/en active Pending
- 2019-12-30 KR KR1020217024220A patent/KR102615834B1/en active IP Right Grant
-
2023
- 2023-04-03 JP JP2023060386A patent/JP2023093517A/en active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050196828A1 (en) * | 2002-08-27 | 2005-09-08 | Zheng Ming H. | Bioreactor with expandable surface area for culturing cells |
US20060134790A1 (en) * | 2003-01-13 | 2006-06-22 | Yasunobu Tanaka | Solid surface with immobilized degradable cationic polymer for transfecting eukaryotic cells |
US20110027889A1 (en) * | 2009-07-28 | 2011-02-03 | Mccarthy Kevin Robert | Synthetic Microcarriers for Culturing Cells |
US20160178491A1 (en) * | 2014-12-22 | 2016-06-23 | Saint-Gobain Performance Plastics Corporation | Capture system of cells and methods |
WO2017169259A1 (en) * | 2016-03-28 | 2017-10-05 | ソニー株式会社 | Cell culture container, cell culture system, cell culture kit and cell culture method |
Non-Patent Citations (1)
Title |
---|
See also references of EP3906294A4 * |
Also Published As
Publication number | Publication date |
---|---|
CN113227344A (en) | 2021-08-06 |
EP3906294A4 (en) | 2022-11-02 |
KR20210097835A (en) | 2021-08-09 |
KR102615834B1 (en) | 2023-12-21 |
JP2023093517A (en) | 2023-07-04 |
JP2022517554A (en) | 2022-03-09 |
EP3906294A1 (en) | 2021-11-10 |
US20200209233A1 (en) | 2020-07-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP2023093517A (en) | Containers storing degradable carriers | |
US5939314A (en) | Distributed pore chemistry in porous organic polymers in tissue culture flasks | |
AU768737B2 (en) | Textured and porous silicone rubber | |
Ariga et al. | Enzyme-encapsulated layer-by-layer assemblies: current status and challenges toward ultimate nanodevices | |
JP5257355B2 (en) | Cell sheet having dimensions, manufacturing method thereof, and cell culture carrier therefor | |
Abdollahi | Extracellular vesicles from organoids and 3D culture systems | |
Pan et al. | Effects of functionalization of PLGA‐[Asp‐PEG] n copolymer surfaces with Arg‐Gly‐Asp peptides, hydroxyapatite nanoparticles, and BMP‐2‐derived peptides on cell behavior in vitro | |
US9440004B2 (en) | Method for preparing biological tissue | |
JP2003534788A (en) | Incubator | |
WO2014039938A1 (en) | Spherical multicellular aggregates with endogenous extracellular matrix | |
EP0806959A1 (en) | Method for preparing macrophages, and kits and compositions therefor | |
WO2017082024A1 (en) | Cell support composite, and method for producing cell support composite | |
EP0402272B1 (en) | Cell-culturing substrate, cell-culturing substrate unit bioreactor and extracorporeal circulation type therapeutic apparatus | |
Zhang et al. | Quantitative collection and enzymatic activity of glucose oxidase nanotubes fabricated by templated layer-by-layer assembly | |
Stanzione et al. | Innovative surface bio-functionalization by fungal hydrophobins and their engineered variants | |
US10745659B2 (en) | Culture container and method for manufacturing culture container | |
US20210115376A1 (en) | Culture container base material, culture container, and production method of culture container base material | |
WO2017006694A1 (en) | Cell culture vessel, cell culture apparatus, and separation/collection method | |
US20150247118A1 (en) | Method for producing 3d cell culture | |
US20200207539A1 (en) | Containers Containing Degradable Carriers | |
JP6690119B2 (en) | Cell culture container and method for producing cell stack | |
JP2018057321A (en) | Cell culture vessel, cell culture system, cell culture method, and method of producing cell culture vessel | |
JP2007308439A (en) | Material suitable for cultivation of floating cell | |
JPH05276923A (en) | Cell culture substrate and its production | |
Labus et al. | Postimmobilization treatments before applications |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 19907890 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2021538428 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 20217024220 Country of ref document: KR Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 2019907890 Country of ref document: EP Effective date: 20210802 |