WO2024059251A1 - Procédé et dispositif support pour permettre l'échange oxygène-dioxyde de carbone - Google Patents

Procédé et dispositif support pour permettre l'échange oxygène-dioxyde de carbone Download PDF

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WO2024059251A1
WO2024059251A1 PCT/US2023/032846 US2023032846W WO2024059251A1 WO 2024059251 A1 WO2024059251 A1 WO 2024059251A1 US 2023032846 W US2023032846 W US 2023032846W WO 2024059251 A1 WO2024059251 A1 WO 2024059251A1
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Prior art keywords
scaffold device
scaffold
carbon dioxide
oxygen
infusion
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PCT/US2023/032846
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English (en)
Inventor
Matthew PHANEUF
Rayan KASSAB
Lisa Fitzgerald
Patrick Hayden
Jayashree CHAKRAVARTY
Nathaniel LONG
Kyle Golden
Arthur Martin
Glenn Gaudette
Richard THYDEN
Luke PERREAULT
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Biosurfaces, Inc.
Boston College
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Publication of WO2024059251A1 publication Critical patent/WO2024059251A1/fr

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    • 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
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/02—Photobioreactors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
    • B01D53/84—Biological processes
    • 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
    • C12M25/02—Membranes; Filters
    • 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
    • C12M43/00—Combinations of bioreactors or fermenters with other apparatus
    • C12M43/04—Bioreactors or fermenters combined with combustion devices or plants, e.g. for carbon dioxide removal

Definitions

  • Cellular agriculture is a process by which animal food products are produced using cells harvested from the animal and grown in cell culture rather than directly from the animal’s tissue, as would be acquired by killing the animal.
  • This process uses tissue engineering techniques to manufacture products, such as meat and dairy products, which are identical to those produced directly from animals.
  • the ability to produce animal-derived products increases food security by eliminating the need for hunting, while also avoiding the environmental impact produced by animal husbandry.
  • scaling up traditional cell culture methods used to grow tissue cultures for this use can be cost prohibitive.
  • tissue cultures often require some form of support or scaffold on which to expand cell growth, as well as a system to remove waste products, such as carbon dioxide and/or lactic acid, from the cells while also enabling a continuous infusion of oxygen.
  • a method of scavenging carbon dioxide and/or metabolic waste and producing oxygen and/or metabolic products comprising: forming a scaffold device from at least one layer of a nano to microporous material, the scaffold device having an interior chamber; providing an infusion into the device comprising at least one of type of whole cells, cellular components or organic material(s); delivering the infusion to the interior chamber of the scaffold device; contacting the scaffold device having the delivered infusion with a solution such as tissue culture medium; and applying sufficient light energy to the device with the infusion in order to initiate oxygen production and carbon dioxide uptake from the surrounding solution.
  • the cellular material includes chloroplasts and/or algae, but alternatively any cell, cellular component or organic materials) can be utilized that can scavenge carbon dioxide and/or metabolic waste and produce oxygen and/or metabolic products and therefore be used in a continuous oxygen-carbon dioxide exchange vis a vis the tissue culture.
  • a method of manufacturing an exchanger for scavenging carbon dioxide and/or metabolic waste and producing oxygen and or metabolic products comprising: forming a nano-microporous porous material; forming a three-dimensional scaffold device from the nano-microporous material, the scaffold device having an interior chamber and an exterior surface; loading an infusion into the interior chamber, wherein the infusion comprises at least one of type of whole cells, cellular components or organic materials); and sealing the interior chamber, wherein the scaffold device is configured for contacting with a tissue culture and enabling light energy applied to the tissue culture to initiate oxygen production and carbon dioxide uptake as an exchange.
  • the sealing can be accomplished using various methods such, but not limited to, ultrasonic welding laser welding and heat cutting
  • the cellular material includes chloroplasts and'or algae, but can alternatively any material can be utilized that can scavenge carbon dioxide and produce oxygen and therefore be used in a continuous oxygen-carbon dioxide exchange vis a vis the tissue culture.
  • an oxygen-carbon dioxide exchanger defined by one or more three-dimensional scaffold devices, each scaffold device being formed from a nano to microporous material and having an outer surface and an interior chamber, wherein the interior chamber of the one or more scaffold devices is sealed at respective edges in order to retain an infusion comprised at least one type of whole cells, cellular components or organic materials, the exchanger being configured such that light energy imparted to the scaffold device creates a renewable oxygen-carbon dioxide exchange.
  • one or more scaffold devices are disposed within a bioreactor or similar vessel.
  • one or more scaffold devices are disposed in relation to a spinner disposed within the vessel.
  • two or more scaffold devices can be disposed within a housing of the vessel, the latter having inlet and outlet ports to add additional components (i.e., cells, cellular components, organic materials) into the scaffold device(s).
  • An advantage realized by the present invention is that a renewable carbon dioxide/oxygen exchange is created, which is especially useful in cellular architecture applications, wherein faster development of large-scale cultures can be obtained and at a lower cost than other already known systems or methods. Additionally, the exchangers created using the herein described method can be scalable.
  • organic materials such as, but not limited to proteins and enzymes, can be bonded or otherwise immobilized to the surface of the scaffold device such as proteins, therapeutic agents, gene sequences and the like, creating additional functionality and versatility, such as lactic acid scavenging or conversion.
  • the herein described device can be made, according to one or more versions, to include a sealable port that is configured to allow initial infusion of target component(s) and refilling, if necessary.
  • the wall of the scaffold device can also include a polyester filter with a pre-set porosity, preferably ranging between 0.1 ⁇ m ⁇ 10 ⁇ m to prevent cells or other components from permeating to the interior of the device.
  • FIG. 1 illustrates an exemplary method of performing a continuous oxygen-carbon dioxide exchange in accordance with aspects of the invention
  • FIG. 2 is a pictorial representation of the method of FIG. 1;
  • FIG. 3 is a further depiction of the method of FIGS. 1 and 2;
  • FIG. 4A illustrates a magnified view of an embodiment of a natural scaffold formed within the human body;
  • FIG. 4B illustrates a magnified view of an embodiment of an electrospun material used to form a scaffold device in accordance with aspects of the invention
  • FIG. 4C illustrates a magnified view of an embodiment of a fabricated scaffold material typically used in industry
  • FIG. 5 is a top view of an embodiment of a scaffold device formed from the electrospun material of FIG. 4B:
  • FIG. 6 illustrates a side elevational view of an embodiment of a bioreactor that can be used in accordance with aspects of the present invention
  • FIG. 7A illustrates the use of the scaffold device of FIG. 5 used for oxygen-carbon dioxide exchange in small cell cultures
  • FIG. 7B illustrates the use of the electrospun material of FIG. 4B to form a plurality of tube-shaped scaffold devices according to aspects of the present invention
  • FIG. 7C illustrates a plurality of scaffold devices fixed to a stirring device to be used to agitate a large cell culture in a bioreactor
  • FIG. 8 illustrates an exemplary embodiment of an oxygen-carbon dioxide exchanging device using an embodiment of the scaffold device made in accordance with aspects of the present invention
  • FIG. 9 illustrates another embodiment of an oxygen-carbon dioxide exchanging device using an exemplary embodiment of the scaffold device.
  • FIG. 10 illustrates another exemplary embodiment of an oxygen-carbon dioxide exchanging device using an embodiment of the scaffold device.
  • the present disclosure generally relates to the growing of a culture of cells in which oxygen is required, such as in the fields of cellular agriculture, mammalian cell culture and biopharmaceuticals. More particularly, in one example, a nanofibrous-microfibrous scaffold is fabricated to surround and contain a biologically active component, which promotes cell growth and culture maintenance. Disclosed herein is a unique method to be used for growing tissue cultures as would be done, for example, in the field of cellular agriculture. However, many or all of the concepts disclosed herein can be applied to other fields such as disease treatment, wound healing, and other such endeavors in which the delivery of a biologically active component using a biocompatible scaffold would be advantageous.
  • FIG. 1 generally illustrates a method 100 for oxygen-carbon dioxide exchange 100 in a tissue cell, which is described in accordance with the present invention.
  • a three-dimensional scaffold device 220 is prepared from a biocompatible material comprising a plurality of nano and micropores, which enable the passage of gas molecules therethrough. While the herein described embodiment focuses on the use of an electrospun material 200 as the biocompatible material, other embodiments may use different biocompatible materials, such as dialysis tubing, in lieu of the electrospun material 200, or other materials may be combined together with the electrospun material 200.
  • the scaffold device 220 is formed into a suitable configuration/shape that further defines an interior chamber, as described in greater detail below
  • an infusion is prepared that is comprised of whole cells and/or cellular components.
  • the infusion is produced using plant tissue cells and includes a chloroplast preparation.
  • the infusion could include algae, isolated chloroplasts or other organic materials capable of scavenging carbon dioxide and'or metabolic waste in order to produce oxygen and/or metabolic produces).
  • the chloroplast preparation according to this embodiment comprises chloroplasts 105 that are present as part of whole plant cells and/or may be separated from or otherwise removed from the whole plant cells using any known method such as, but not limited to heating leaves in water and then placing in alcohol or grinding chilled leaves in solution and then filtering chloroplasts. Examples are further described in Spencer, D., & Unt, H. (1965). Biochemical and Structural Correlations in Isolated Spinach Chloroplasts Under Isotonic and Hypotonic Conditions. Australian Journal Of Biological Sciences. 18(2), 197; Jensen, R., & Bassham, J. (1966). Photosynthesis by isolated chloroplasts. Proceedings Of The NationalAcademy Of Sciences, 56(4), 1095-1101; and Nass, M.
  • the infusion is then delivered or infused into the interior chamber of the scaffold device 220.
  • This step 106 of the method 100 may further include sealing the infusion within the formed interior chamber of the scaffold device 220.
  • the scaffold device 220 loaded with the infusion is placed in contact with a solution such as a tissue cell culture medium, such as, for example, a muscle tissue cell culture, wherein the contact occurs within a bioreactor or other such vessel 300 (FIG. 5), as described in greater detail in a later section of this application.
  • a tissue cell culture medium such as, for example, a muscle tissue cell culture
  • Sufficient light energy 250 is then applied at step 110 to the scaffold device(s) in order to initiate an oxygen-carbon dioxide exchange.
  • the light energy 250 can be applied at a different point in the method, such as prior to step 106.
  • the light energy 250 is converted to chemical energy by the contained and sealed chloroplasts 105, while freeing up oxygen from the surrounding water that is present in the culture medium
  • the chemical energy according to this embodiment is in the form of adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide phosphate (NADPH).
  • ATP adenosine triphosphate
  • NADPH nicotinamide adenine dinucleotide phosphate
  • the ATP and NADPH convert carbon dioxide present in the tissue culture as a waste product into glucose in accordance with the Calvin cycle.
  • oxygen that is produced by the chloroplasts 105 is released into the tissue culture and used by the cells therein, while excess carbon dioxide produced by the cells in the tissue
  • an efficient and continuous oxygen-carbon dioxide exchange can be produced using a biocompatible scaffbld/scaffbld device(s) and a renewal biologically active component.
  • This efficient and continuous oxygen-carbon dioxide exchange enables faster development of large tissue cultures at a lower cost.
  • This method is shown pictorially in FIG. 2, showing the chloroplast as detailed more specifically by its individual components as labeled, which is used as the basis of the remainder of the infusion (not shown in this view) delivered to the scaffold device having a sealable interior chamber.
  • light energy delivered at a specific wavelength or range of suitable range of wavelengths over a defined duration of time and following contact with a tissue (e.g., muscle) sample creates the renewable carbon dioxide exchange and developments of the contacted tissue.
  • FIG. 3 depicts another pictorial representation of a more specific embodiment of the herein described method in which the surface of a scaffold device used in an exchanger or exchange device is chemically-modified to create functional groups, such as described in Bide et al. US Patent No. 7,037,527B2 that are used to ionically or covalently attach biologic molecules such as, but not limited to, proteins, glycoproteins, synthetic compounds, or oligonucleotides that can provide an additional targeted effect to either the outer or inner surface of the device.
  • an enzyme such as lactate dehydrogenase could be covalently attached to the functional groups on the surface.
  • a cell chamber which is the interior chamber of the scaffold device 220 is created wherein the defined chamber is formed and sterilized by known means (chemically, ultraviolet irradiation, electron beam, etc.) per step 302.
  • the chloroplast preparation/infusion is also sterilized prior to delivery per step 304, also using known techniques.
  • the chloroplast preparation/infusion is then introduced for delivery into the defined chamber according to step 306 and a number of the scaffold devices 220 each having the delivered infusion are further introduced into a bioreactor (not shown in this view) or similar exchanger vessel or housing per step 308.
  • the sealed chambers) of the exchanger are then exposed to sufficient light energy for a predetermined time and wavelength to initiate the carbon dioxide'oxygen exchange per step 310.
  • At least a portion of the scaffold device 220 is produced using electrospinning technology. Aspects of such electrospinning technology are further described in U.S. Patent Nos. 7,413,575. 8,771,582, 10,328,032, and 10,441,550, as well as U.S. Patent Application Publication No. 2006/0200232 Al), the contents of each of which are hereby incorporated by reference in their entirety. The relevant aspects of this methodology are herein described.
  • the electrospinning process is done at room temperature (about 25°C) and can therefore, easily incorporate drugs or bioactive agents (such as gene sequences) directly into the nanofibers of the electrospun materials used in the scaffold device. This is advantageous because many existing electrospun materials are produced at high temperatures, which makes it impractical to incorporate heat-sensitive protein-based drugs or bioactive agents into the electrospun material during manufacturing.
  • a method of forming an electrospun material 200, FIG. 4B, for purposes of this embodiment comprises dissolving a non-biodegradable polymer in an organic solvent to produce a polymer solution.
  • the polymer solution is loaded into an electrospinning instrument configured to be set at a specified flow rate.
  • An electrical current of 15-30 kV is applied to a needle of the electrospinning instrument and the needle is placed at a predetermined distance, preferably 5cm- 50cm from a collecting surface.
  • the polymer solution is electrospun onto the collecting surface for a period of time to form an electrospun polymer material. As shown in FIG. 3, the electrospun material 200, FIG.
  • a scaffold device 220 for purposes of forming a scaffold device 220 may be formed in a series of layers stacked on top of each other, wherein each layer can be formed from a different polymer solution.
  • one layer 224 of an exemplary scaffold device 220 may be formed using nPU and a subsequent layer 232 may be formed from riPET-PBT with a PET filter layer 236 being positioned between the layers 224 236 also as shown in the cross section provided at FIG. 3.
  • the layers 224, 232 can be reversed in relation to the intermediate PET filter layer 236.
  • a single polymer material can be utilized for each of the layers surrounding the intermediate filter layer 236.
  • two or more polymers could be electrospun and formed in layers, but without the use of the intermediate PET filter layer.
  • polymer or polymers used in addition to other aspects of the electrospinning process enable control of the porosity of the finished electrospun material 200, FIG. 4B.
  • a plurality of micropores or nanopores are introduced into the electrospun material 200 during the electrospinning process.
  • one or more drugs and/or bioactive agents can be incorporated into one layer or alternatively each layer in order to create a bioreactive surface; for example, attachment of lactate dehydrogenase to convert lactic acid to pyruvate.
  • one or more binding factors are incorporated into the electrospun material 200, FIG 4B, (or other biocompatible material) to aid in binding the electrospun material 200, FIG. 4B, to target molecules, wherein the surface of the herein formed scaffold device can be suitably modified in order to create functional groups for ionic interactions.
  • FIG. 4A illustrates a magnified view of an example of a scaffold 400 that is natural to a human body.
  • the scaffold 400 can be comprised of a plurality of collagen strands 404 and generally supports body tissue, such as muscle tissue or the tissue of internal viscera.
  • body tissue such as muscle tissue or the tissue of internal viscera.
  • the natural scaffold 400 has a plurality of nanopores 406, which are distributed throughout as shown that enable small molecules or even single cells to move through the natural scaffold 404.
  • FIG. 4B illustrates a similarly magnified view of a disclosed electrospun material 200 that is used to fabricate an embodiment of the herein described scaffold device 220
  • the individual nanofibers 204 of the electrospun material 200 are clearly discernable and comprise a web-like pattern.
  • a plurality of nanopores 208 are defined between the individual nanofibers 204 that may enable small molecules or even single cells to move through the electrospun material 200.
  • the structural similarity of the electrospun material 200 and the natural scaffold 400 can clearly be seen when comparing FIG. 4A and FIG. 4B.
  • FIG. 4C illustrates a similarly magnified view of an example of a fabricated scaffold 500 that is currently used in industry.
  • the nanofibers 504 of this specific scaffold 500 are much larger than the nanofibers 204 of the electrospun material of FIG. 4B or the plurality of collagen strands 404 of the natural scaffold 400, previously shown according to FIG. 4A.
  • the nanofibers 504 of the scaffold 500 are aligned in a more regular pattern of approximately parallel nanofibers. Accordingly, the spaces or pores between the nanofibers of FIG. 4C are larger than those of the natural scaffold 400 in FIG. 4A, or the electrospun material 200 of FIG. 4B.
  • the residual organic solvent is removed from the electrospun material 200 and the electrospun material 200 is removed from the target surface.
  • the finished electrospun material 200 can then be formed into a three-dimensional scaffold device 220, FIG. 5, having a defined interior chamber made up of one or more layers, such as shown in FIG. 3. Aspects of forming the scaffold device 220 are discussed in PCT Application WO2021/188814 Al (International publication no. PCT/US2021/022994), the contents of which are herein incorporated by reference. Referring to FIG.
  • the electrospun material 200 may be cut to the desired shape and size and the edges of the electrospun material 200 may be partially or fully sealed to form a three-dimensional scaffold device 220, defining an interior chamber and having a cross section like or similar to that shown in FIG. 3. Sealing of the edges 210, FIG. 5, of the scaffold device 220 can be done using any method known in the art.
  • the scaffold device 220 is formed in a “frameless” manner in which no additional structural component is used or required in order to form the scaffold device 220.
  • other embodiments of the scaffold device 220 may be attached to a frame, the latter of which remains part of the finished scaffold device 220.
  • an exterior surface of the scaffold device 220 be comprised from a first polymer solution and an inside surface be comprised from a second polymer solution as shown, for example, according to the device 220 version shown in FIG. 3.
  • the scaffold device can be edible or biodegradable. In this manner, it may not be necessary to remove the scaffold device 220 from its application.
  • One or more surfaces of the scaffold device 220 may be further modified to create functional groups tor ionic interaction to attract and promote attachment of certain proteins/cells/cellular components. As shown in FIGS.
  • the scaffold device 425, 525 may not be configured to contain cells or cell components and may be used to separate cells or a cell culture from the chloroplast preparation. Jn these latter embodiments, the scaffold device 425, 525 having the electrospun material 200, which can be microporous or nanoporous, acts as an interface through which oxygen-carbon dioxide exchange occurs.
  • the wall of the scaffold device 220 can also include a polyester filter with a pre-set porosity, preferably ranging between 0.1 ⁇ m and 10 ⁇ m, to prevent cells or other components from permeating to the interior of the scaffold device 220.
  • chloroplasts 105 FIG. 2, which are specialized organelles present in the cells of plants and green algae.
  • the chloroplasts 105 may be obtained and isolated from plant material using an existing isolation procedure.
  • the chloroplasts 105 may then undergo one or more post-isolation processing steps to form a chloroplast preparation for infusion into the formed interior chamber of the scaffold device 220.
  • a chloroplast preparation can be made using whole plant cells such that the oxygen-carbon dioxide exchange properties of the chloroplasts 105 may be utilized without requiring an isolation step. Referring to FIG.
  • the scaffold device 220 may be fitted with a port 202 that is fluidly connected to the defined interior chamber in order to aid in the infusion of a chloroplast or similar preparation into the scaffold device 220. Once the infusion preparation has been delivered or otherwise loaded into the interior chamber of the scaffold device 220 the port 202 may be closed or otherwise sealed [0046]
  • the loaded scaffold device 220, FIG. 5, is then introduced into the cell culture, which may be grown in a bioreactor or other suitable vessel 600, FIG. 6, as discussed herein.
  • the scaffold device 220 can be used as oxygen-carbon dioxide exchangers in smaller cell cultures which are grown in flasks or beakers 60, or even well plates 50.
  • FIG. 7 A the scaffold device 220 can be used as oxygen-carbon dioxide exchangers in smaller cell cultures which are grown in flasks or beakers 60, or even well plates 50.
  • FIG. 7 A the scaffold device 220 can be used as oxygen-carbon dioxide exchangers in smaller cell cultures which are grown in flasks or beakers 60
  • each scaffold device 220 may comprise one or more lengths of tube 224 that contain the chloroplast preparation and can be directly added to the cell culture or are used to cycle chloroplast preparation through the cell culture, the latter of which is disposed in various levels of a cylindrically shaped bioreactor vessel 300.
  • one or more loaded scaffold devices 220 may be attached to a rotating shaft component 704 of a stirring device 700, the latter being disposed, for example, within a bioreactor vessel such as a vessel 300.
  • the stirring device 700 is configured to rotate the one or more loaded scaffold devices 220 relative to the vessel 300 to promote movement of carbon dioxide molecules into the interior chamber of each scaffold device 220 via the micro/nanopores and of oxygen molecules from the defined interior chamber via the micro/nanopores.
  • a source of light energy 250, FIG. 2 such as light-emitting diode(s) (LED), compact fluorescent (CFL), incandescent, and/or halogen light sources can be emitted at a particular wavelength and duration, for application either prior to putting cells into the bioreactor vessel or after cell placement in the vessel 300, which “activates” the oxygen-carbon dioxide exchange process.
  • the amount of light energy 250 and the duration of the application of light energy 250, FIG. 2 will vary depending on factors such as, but not limited to culture temperature, culture type and the ratio of culture to chloroplast preparation.
  • the scaffold device 220 may further act as a support for the cells of the tissue culture. In this manner, the cells of the tissue culture can attach to the exterior of the scaffold device 220 and grow while the interior of the scaffold device 220 can hold or otherwise support a different type of cell or material.
  • Other embodiments of the scaffold device 220 may be impregnated with a bioactive agent to further aid in the growth of the cell culture and/or recycle the media. For example, when the tissue culture is comprised of muscle cells, a build-up of lactic acid will then occur as the muscle cells breakdown carbohydrates for energy when oxygen levels are low. To alleviate this issue, lactate dehydrogenase can be incorporated into the electrospun material 200, FIG. 4B, or otherwise attached to the scaffold device(s) 220 to convert the lactic acid to pyruvate.
  • an embodiment of an oxygen-carbon dioxide exchanger 320 is shown that includes a housing 322 that surrounds one or preferably a plurality of scaffold device(s), such as device 220, each comprising a nano-microporous material either created by an electrospinning process or provided by tubing, as previously discussed.
  • the housing 322 defines an inlet port 324 and a corresponding outlet port 326 for a chloroplast preparation to be passed through the interior of the housing 322.
  • the housing 322 further defines separate media inlet and outlet ports 327, 329 for passing media containing cells or cell culture through the interior of the housing 322.
  • the nano-microporous material of the scaffold device(s) 220 located within the interior of the housing 322 acts to separate the media and the chloroplast preparation inside of the housing 322.
  • Each scaffold device acts as an interface enabling an oxygen-carbon dioxide exchange between the chloroplast preparation and the media. In this manner, excess carbon dioxide is removed from the media and exchanged with excess oxygen from the chloroplast preparation.
  • the media and the chloroplast preparation flow in opposite directions relative to each other per the respective ports of the housing 322.
  • FIG. 9 illustrates another embodiment of an oxygen-carbon dioxide exchanger 420 that includes a plate-like housing 422, made from a suitable structural and biocompatible material that surrounds a scaffold device 425 comprising a nano-microporous material, such as an electrospun material as previously discussed.
  • the housing 422 defines an inlet port 424 and a corresponding outlet port 426 for a chloroplast preparation to be passed through the interior of the housing 422.
  • the housing 422 further defines first and second media inlets 423, 427 and corresponding first and second outlet ports 428, 429 for passing media containing cells or cell culture through the interior of the housing 422
  • the scaffold device 425 retained within the interior of the housing 422 of the exchanger 420 acts to separate the media and the chloroplast preparation inside of the housing 422.
  • the retained scaffold device 425 further acts as an interface enabling an oxygen-carbon dioxide exchange between the chloroplast preparation and the media. In this maimer, excess carbon dioxide is removed from the media and exchanged with excess oxygen from the chloroplast preparation.
  • the media and the chloroplast preparation flow in opposite directions relative to each other based on the ports provided on the housing 422.
  • FIG. 10 illustrates yet another embodiment of an oxygen-carbon dioxide exchanger 520 that includes a first plate 522 comprising a first plurality of wells 523 and a second plate 532 comprising a second plurality of wells 533.
  • Each of the first plurality of wells 523 includes a top opening 523a and a bottom opening (not shown).
  • Each of the second plurality of wells 533 includes a top opening 533a.
  • a scaffold device 525 comprised of a nano-microporous material, such as an electrospun material, is positioned between the bottom openings (not shown) of the first plurality of wells 523 and the top openings 533a of the second plurality of wells 533.
  • a chloroplast preparation is deposited in each of the first or second plurality of wells 523, 533 and a media containing cells or cell culture is deposited in each of the remaining wells.
  • the position of the scaffold device 525 prevents mixing of the chloroplast preparation and the media, but acts as an interface which enables excess carbon dioxide to be removed from the media and exchanged with excess oxygen from the chloroplast preparation in a renewable manner.
  • the disclosure discusses a specific embodiments of a scaffold device being used for oxygen-carbon dioxide exchange and more specifically for applications involving cellular architecture, it should be realized that the scaffold device can be tailored to suit a variety of other uses.
  • the scaffold device can be infused with target cells, such as chloroplasts and placed or applied to a living body to promote specialized tissue growth and/or repair such as, for example, delivering oxygen and/or a bioactive agent to a wound.
  • a method or device that “comprises,” “has.” “includes,” or “contains” one or more steps or elements possesses those one or more steps or elements, but is not limited to possessing only those one or more steps or elements.
  • a step of a method or an element of a device that “comprises,” “has,” “includes,” or “contains” one or more features possesses those one or more features, but is not limited to possessing only those one or more features.
  • a device or structure that is configured in a certain way is configured in at least that way, but may also be configured in ways that are not listed.
  • Additional embodiments include any one of the embodiments described above and described in any and all exhibits and other materials submitted herewith, where one or more of its components, functionalities or structures is interchanged with, replaced by or augmented by one or more of the components, functionalities or structures of a different embodiment described above.
  • scaffold device

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Abstract

Procédé de piégeage du dioxyde de carbone et/ou des déchets métaboliques et de production d'oxygène et/ou de produits métaboliques consistant à constituer un dispositif support à partir d'un matériau nanométrique ou microporeux de manière à ce que le dispositif support délimite une chambre interne. Une perfusion contenant au moins un type de cellule entière, des composants cellulaires contenant des chloroplastes ou des matières organiques est mise à disposition. La perfusion est acheminée dans la chambre interne du dispositif support et le dispositif support chargé de l'infusion est mis en contact avec une culture tissulaire. De l'énergie lumineuse est appliquée à la culture tissulaire à une longueur d'onde et une durée particulières afin d'initier la production d'oxygène et l'absorption de dioxyde de carbone par les chloroplastes de manière renouvelable. Cette méthode permet de mettre au point des échangeurs d'oxygène-dioxyde de carbone variés et évolutifs. En outre, au moins un agent bio-réactif peut être immobilisé sur les nano-matériaux microporeux du dispositif support.
PCT/US2023/032846 2022-09-15 2023-09-15 Procédé et dispositif support pour permettre l'échange oxygène-dioxyde de carbone WO2024059251A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070231887A1 (en) * 2006-03-14 2007-10-04 University Of Rochester Cell culture devices having ultrathin porous membrane and uses thereof
US20100297768A1 (en) * 2003-11-05 2010-11-25 Michigan State University Nanofibrillar structure and applications including cell and tissue culture
WO2018235745A1 (fr) * 2017-06-20 2018-12-27 日本毛織株式会社 Tissu non tissé à fibres longues biocompatible, son procédé de production, échafaudage tridimensionnel pour culture cellulaire, et procédé de culture cellulaire l'utilisant
US20190316067A1 (en) * 2016-12-01 2019-10-17 Arborea Ltd Photo-bioreactor device and methods

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100297768A1 (en) * 2003-11-05 2010-11-25 Michigan State University Nanofibrillar structure and applications including cell and tissue culture
US20070231887A1 (en) * 2006-03-14 2007-10-04 University Of Rochester Cell culture devices having ultrathin porous membrane and uses thereof
US20190316067A1 (en) * 2016-12-01 2019-10-17 Arborea Ltd Photo-bioreactor device and methods
WO2018235745A1 (fr) * 2017-06-20 2018-12-27 日本毛織株式会社 Tissu non tissé à fibres longues biocompatible, son procédé de production, échafaudage tridimensionnel pour culture cellulaire, et procédé de culture cellulaire l'utilisant

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