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WO1997016527A1 - Materiel de culture pour foie bio-artificiel - Google Patents

Materiel de culture pour foie bio-artificiel Download PDF

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Publication number
WO1997016527A1
WO1997016527A1 PCT/US1996/017559 US9617559W WO9716527A1 WO 1997016527 A1 WO1997016527 A1 WO 1997016527A1 US 9617559 W US9617559 W US 9617559W WO 9716527 A1 WO9716527 A1 WO 9716527A1
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WO
WIPO (PCT)
Prior art keywords
blood
bioreactor
hemoperfusion
circuit
media
Prior art date
Application number
PCT/US1996/017559
Other languages
English (en)
Inventor
Darrell Page
Robert Wojciechowski
Original Assignee
Cellex Biosciences, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cellex Biosciences, Inc. filed Critical Cellex Biosciences, Inc.
Priority to JP9517579A priority Critical patent/JPH10512159A/ja
Priority to EP96938724A priority patent/EP0801674A4/fr
Publication of WO1997016527A1 publication Critical patent/WO1997016527A1/fr

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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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/10—Hollow fibers or tubes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/022—Artificial gland structures using bioreactors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/14—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
    • A61M1/16—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis with membranes
    • A61M1/1621—Constructional aspects thereof
    • A61M1/1623—Disposition or location of membranes relative to fluids
    • A61M1/1625—Dialyser of the outside perfusion type, i.e. blood flow outside hollow membrane fibres or tubes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
    • A61M1/3472—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate
    • A61M1/3475—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate with filtrate treatment agent in the same enclosure as the membrane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
    • A61M1/3472—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate
    • A61M1/3486—Biological, chemical treatment, e.g. chemical precipitation; treatment by absorbents
    • A61M1/3489—Biological, chemical treatment, e.g. chemical precipitation; treatment by absorbents by biological cells, e.g. bioreactor
    • 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
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • 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
    • C12M23/16—Microfluidic devices; Capillary tubes
    • 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
    • C12M23/28—Constructional details, e.g. recesses, hinges disposable or single use
    • 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
    • C12M23/58—Reaction vessels connected in series or in parallel
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/10—Perfusion
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2205/00—General characteristics of the apparatus
    • A61M2205/75—General characteristics of the apparatus with filters
    • A61M2205/7554—General characteristics of the apparatus with filters with means for unclogging or regenerating filters

Definitions

  • the present invention relates to methods and systems for bioartificial liver support.
  • the invention relates to cultureware useful for supporting the function of hollow fiber bioreactors.
  • the invention relates to dialysis and patient support systems, and particularly those that employ means for exchanging gases in the blood or other fluids.
  • C3A cloned liver cell line
  • hepatocytes are suspended in a collagen solution and inoculated into the lumen of a hollow fiber assembly.
  • the cell/collagen solution is then incubated with media for approximately 20 hours under conditions that promote collagen gel contraction. This results in the formation of a biomatrix containing the collagen and cells, with the formation of an open intraluminal space within the fibers.
  • blood is allowed to flow through the extraluminal shell space, with the semi-permeable hollow fibers serving to separate the blood from the hepatocyte-containing matrix.
  • An intraluminal stream of medium is allowed to flow through the newly created lumen space, to carry the growth factors and nutrients necessary to support cell growth.
  • the intraluminal stream can also be used to remove toxins and metabolic products.
  • the Sielaff et al. article provides a functional diagram of the cell culture hardware and software used in regulating pH, oxygen, and shell flow at both the prehemoperfusion and hemoperfusion phases.
  • the diagram depicts the use of a gas exchanger ("GEX") to provide a blend of air and CO 2 .
  • GEX gas exchanger
  • Gas exchangers were developed and continue to be used for such purposes as open heart surgery, where the patient's own heart and lungs are shut down during surgery.
  • the function of such organs is primarily mechanical, as compared to the function of the liver, which is tremendously complex and primarily metabolic.
  • Figure 1 shows a disposable cultureware assembly of the present invention, in combination with two bioreactors, in operation in the course of the culture circuit;
  • Figure 2 shows the assembly and bioreactors of Figure 1 in operation in the course of the hemoperfusion circuit
  • Figure 3 shows a perspective view of a preferred switching valve for use in an apparatus of the present invention
  • Figures 4 A and 4B show, respectively, front and rear perspective views of an operable hemoperfusion system incorporating the assembly and bioreactors of Figures 1 and 2.
  • the present invention provides a disposable cultureware (i.e., bioreactor support module and tube set) assembly for use with a bioartificial liver employing a hollow fiber bioreactor having an extracapillary flow path and an intraluminal flow path, the assembly comprising: a) a culture circuit for delivering media to the extracapillary flow path of a bioreactor; b) a hemoperfusion circuit for delivering blood to the extracapillary flow path of a bioreactor; c) switching means operably connected to both the culture circuit and hemoperfusion circuit, for controlling between the delivery of media and blood to the extracapillary flow path; d) a gas exchange cartridge operably connected to both the culture circuit and the hemoperfusion circuit, for controllably gassing the media and blood prior to delivery to a bioreactor; e) a lumen circuit for delivering fluids to the lumen flow path of a bioreactor; and f) bioreactor support and attachment means for operably connecting the extracapillary and lumen flow paths of a
  • hepatocytes or other suitable cells are grown and maintained within the lumen of the bioreactor hollow fibers, as opposed to the extracapillary space.
  • the collagen/cell combination is able to constrict in order to entrap the cells within a gel-like matrix, thereby opening up a media flow path within the lumen.
  • oxygenated media or blood are delivered to the cell/collagen matrix across the semi-permeable membrane forming the hollow fibers.
  • the invention provides a hollow fiber bioreactor operably connected to a cultureware assembly as described herein.
  • the cultureware of the present invention is used to support one or more bioreactors for operation within a cell culture system that provides for the basic mammalian cell functions required for cell sustainment.
  • a cell culture system that provides for the basic mammalian cell functions required for cell sustainment.
  • Such an instrument typically provides an incubator to ensure a 37°C temperature environment and a circulation system (including pumps, tubing, controls, etc.) to provide the cells with nutrients and oxygen.
  • Its integral gassing system supplies oxygen as an essential nutrient and carbon dioxide for pH control.
  • the system has a control panel for user interface to the processor for pump management and for input of sensor and alarm settings.
  • a customized software program allows for varied instrument set up parameters to accommodate varying patient requirements.
  • the invention provides a complete system comprising a cultureware assembly of the present invention, in combination with the bioreactors, pumps, power supply, and other components for its operation in hemoperfusion.
  • the cultureware is particularly unique in view of its use of a single gas exchange cartridge within both the media and blood circuits.
  • the ability to employ a single cartridge is surprising from the perspective of both engineering and performance functions, and provides significant benefits and options, in terms of production costs, operation and the like.
  • Figures 1 and 2 provide alternative flow diagrams employing a preferred embodiment of the disposable cultureware assembly 10, including both the bioreactor support module 11 and tube set 12.
  • assembly 10 includes two fluid circuits useful for serving the extracapillary flow path of a bioreactor; a culture circuit 14 and a hemoperfusion circuit 16.
  • Culture and hemoperfusion circuits 14 and 16 are each operably connected to switching valve 18. The switching valve can be controlled to select either the culture or hemoperfusion path to be used in operation.
  • Assembly 10 also provides a lumen circuit, for serving the lumen flow path of bioreactor.
  • Figure 3 provides a perspective view of a preferred switching valve for use within an assembly of the present invention. Depicted in Figure 3 is a mechanical valve suitable for manual operation. Those skilled in the art, given the present description, will understand the manner in which automated valve means could similarly be provided, to be operated by control means located, for instance, on the instrument panel.
  • the valve shown in Figure 3 provides a rigid box-like container 70 having walls 72 and 74 supporting a removable cover portion 76 and forming opposite ends 78 and 80.
  • Cover portion 76 provides access to the interior of the container.
  • Walls 72 and 74 are provided with matching inlet 82 and outlet ports 84, provided through the walls to allow pairs of tubes 86 and 88 to enter and exit the container, traversing the interior.
  • the pairs of tubes are part of the media and blood lines, respectively, serving the extracapillary circuit.
  • a shaft 90 is rotatably retained within and substantially through the container, the shaft having a plurality of circumferential grooves 92 and 94 configured to simultaneously open and close each pair of tubes.
  • the grooves are provided in the form of recessed regions each having a cam-like appearance when viewed in Figure 3B along the axis of the shaft.
  • the regions are recessed slightly (i.e., the cam portion having the greater axis), to allow a tube to be retained in a pinched (i.e. , closed) configuration.
  • the regions are recessed more deeply on the opposite side of the shaft, to allow the tube to return to its original dimensions.
  • the walls of the groove are also preferably tapered 96 slightly inward (i.e., toward each other). This enables the walls of the groove to participate in re-opening the tubes by gently pinching the tubes along an axis parallel to the axis they were pinched closed.
  • the shaft is then positioned longitudinally within the container so that, with the cover closed, a tube of appropriate outer diameter can be pinched between the raised cam portion and an interior surface.
  • a pair of tubes serving a circuit can therefore be simultaneously closed by rotating the shaft in such a matter that the raised cam portion pinches the tubes closed against the cover or base.
  • the shaft is rotatably positioned through apertures 98 and 100 in the ends of the container.
  • the shaft extends beyond one end, where it can be attached on the exterior of the container to a handle 102 that can be grasped by the user and rotated, e.g. , 180 degrees to close the previously opened circuit and open the previously closed one.
  • Both the "culture” and “hemoperfusion” fluid paths can be selectively operated by use of the switching valve for circulating fluids to the shell (i,e. , extracapillary) side of the hollow fiber bioreactors. This allows for mass transfer of gases, nutrients and impurities through the semipermeable membrane to the lumen side which contains the hepatocytes.
  • Gas exchange cartridges suitable for use in the assembly of the present invention are available from a number of sources.
  • suitable cartridges include the "Capiox II” brand hollow fiber oxygenator available from Terumo Corporation (Japan), the “Safe-1” brand hollow fiber oxygenator from Polystan A/S (Denmark), the “Maxima Plus” brand oxygenator available from Medtronic Cardiopulmonary (Anaheim, CA), and the "Affinity” brand hollow fiber oxygenator and the ECMO brand membrane oxygenator, both available from
  • an oxygenator in a suitable style and providing an optimal combination of such characteristics as the choice of membrane material, membrane pore size and surface area, fiber inside diameter and wall thickness, capillary lumen volume and extracapillary volume, port placement and dimensions, recommended maximum media or blood flow rate.
  • gas exchange using a hollow fiber oxygenator will be achieved and controlled by adjusting such factors as the gas composition and flow rate and the liquid (media or blood) pressure and flow rate.
  • Media or blood are able to flow through the lumen of the hollow fibers as gas flows across the outside of the lumen (extracapillary space).
  • a preferred system of the present invention also includes means for monitoring the blood gas values in the extracapillary circuit during hemoperfusion. Variations in pO 2 , pCO 2 and pH levels are each useful in determining blood gas values during hemoperfusion.
  • Variations in these parameters can occur due to patient condition, control settings and equipment performance. Variations can be regulated by suitable control of oxygenator function (e.g. , gas composition, flow rate) and other parameters. Intermittent sample techniques can be used, but may miss or mask dramatic fluctuations. As a result, real time status of hemoperfusion management, in the form of continuous monitoring, is preferred.
  • the pH probe can take any suitable form, e.g. , including those that make direct contact with the media or blood, and those that make only indirect contact. Examples of suitable pH probes include the "TH" line of pH electrodes available from Ingold, the "ASI" line of electrodes available from Analytical Sensors, Inc.
  • Electrode Company (Houston, TX), and the "GX" line of electrodes available from pHoenix Electrode Company (Houston, TX).
  • Preferred electrodes provide an optimal combination of such properties as accuracy, safety, storage/regeneration properties, temperature compensation, and compatibility with the patient's blood within the temperature, flow and other conditions of use.
  • Electrodes are available in a number of sizes (e.g., 3mm- 20mm), sensor types (e.g., gold, antimony, platinum, silver), connector types (E.G., U.S.
  • Standard, BNC, pin-type, plug- type electrode types (e.g., metallic, reference, conductivity), and electrode body construction (e.g., PVC, teflon, glass, epoxy/polymer), and shapes (flat surface, dome- shaped, hemispherical, bulb).
  • electrode types e.g., metallic, reference, conductivity
  • electrode body construction e.g., PVC, teflon, glass, epoxy/polymer
  • shapes flat surface, dome- shaped, hemispherical, bulb.
  • the system of the present invention can employ other suitable means to monitor and/or alter the blood gas composition (and pH) in the course of operation.
  • Extracorporeal blood gas monitoring systems such as the "Cardiomet 4000” system available from Biomedical Sensors Limited (England), and the “CDl System 400” system available from 3M Health Care (Valencia, CA) are examples of such systems.
  • the cultureware can be used to form a complete hemoperfusion system 50, including the assembly of the present invention and first and second bioreactors, all housed within instrument cabinet 52. Also seen within the system are blood monitor panel 54, a universal power supply 56, an IV pole 58, pressure gauges 60, a media bag hangar 62, as well as supporting cart 64 itself. As seen from the rear view provided in Figure 4B, the system also includes an on/off switch 66 controlling the bioreactor support module, a printer port 68, an oxygen tank and regulators 70 as well as a CO 2 tank and regulators 72, and a power cord 74.
  • First and second bioreactors can be prepared and used in the manner described in PCT Patent Application No. PCT//US91/07952 , the disclosure of which is incorporated herein by reference.
  • Suitable bioreactors for use in a system of the present invention can be constructed using available materials and methods.
  • a suitable gas blending system is capable of operable attachment to the gas exchange cartridge 20 and preferably includes supplies of medical grade oxygen, medical grade carbon dioxide, and room air gas.
  • the air, oxygen, and carbon dioxide gas flows are controlled and mixed to provide an oxygen-enriched and variable carbon dioxide supply to the gas exchange cartridge.
  • the oxygen is needed to provide additional oxygen to the liver cells by means of oxygenating the media solution during the culture phase and oxygenating the patient's venous blood during the treatment mode.
  • the carbon dioxide gas flow is controlled to modulate the pH of the media during the culture phase and the patient's blood during the hemoperfusion phase.
  • the gas delivery system includes a number of components to ensure proper gas delivery timing and pressures.
  • Such components include, but are not necessarily limited to, the use of in-line pressure relief valves, diaphragm air pumps rated for continuous service, in-line coalescent filters to remove particulate material, and separate precision regulators, e.g., separate precision needle valves, to provide separate control for each gas.
  • a preferred gassing system further includes medical grade oxygen and carbon dioxide gas cylinders, high pressure regulators, and monitoring gauges.
  • the gas cylinders provided are of sufficient size to provide for continuous operation of the gassing system of the instrument for ten days without a cylinder change.
  • containers for fiuid storage including containers for storing extracapillary media, lumen-in media, lumen-out media, and a waste bottle, together with appropriate tubing, valves, holders, and the like.
  • the system also provides several pumps for moving fluids through the system. These are of two types: peristaltic pumps and bellows pump.
  • a blood pump described in greater detail below, is designed to operate on human blood.
  • Other pumps in the system are typically employed for delivery or moving the various solutions (i.e., media, waste, etc.).
  • a bellows pump is preferably used during culture phase to propel media through the shell side of the bioreactors.
  • Peristaltic pumps are intended to deliver quantities of media to the hepatocytes over several days without loss of accuracy.
  • the system employs two bioreactors, with their extracapillary flow paths connected in series (and their lumen flow paths connected in parallel), four of the peristaltic pumps are "ganged" in pairs, a configuration envisioned by the manufacturer of these pumps and their drive motors.
  • the system also provides an incubator, preferably in the form of a forced draft convection system, designed to keep the cultureware at a controlled temperature.
  • a fan on the incubator circulates heated air from a coil heater to provide pre-set ambient air conditions.
  • Inside the incubator are typically rows of bulkhead fittings used for electrical cable interface and gas supply to the cultureware.
  • a variety of blood pump modules are suitable for use in the system of the present invention, in order to control and supervise the extracorporeal blood circuit.
  • a preferred blood pump module will include with the pump, an arterial and venous pressure sensor/alarm, a drip chamber air detector, arterial and venous solenoid clamps for blood line clamping for certain alarms, and an optional heparin pump. Such a module can be used to circulate the blood from the patient to the tube set and return it to the patient during the hemoperfusion phase of the procedure.
  • a suitable uninterruptible power system (“UPS") is used to provide battery backed A.C. line voltage for the bioreactor support and blood monitoring modules; A.C. line power conditioning for the two modules; and A.C. line isolation which brings patient leakage current to an acceptable level.
  • UPS uninterruptible power system
  • the instrument and consumable supplies are preferably installed on a cart to provide a mobile self-contained platform for operation.
  • Assorted docking grids, hangers and holders can be provided, e.g., for hanging media bags and waste containers on the cart, and for docking and securing the blood pump and power supply, and bioreactor support module.
  • the software provides the following functions: 1) user interface via a keypad, display, audio alarm, and printer, 2) pump controls, 3) maintenance of incubator temperature, 4) control of O 2 and CO 2 gassing and maintenance of pH during the culture phase, and 5) provision of timing and sequential control of operations.
  • viable primary hepatocytes can be recovered from pigs and prepared in the manner described in their above-captioned article of Sielaff et al. This article goes on to describe the need for liver support devices to provide a substantial viable hepatocyte mass exhibiting differentiated phenotype, and biotransformation function.
  • the article describes the use of a bioartificial liver based on the cell-entrapment hollow fiber bioreactor of Scholz and Hu. See, for example, Patent Cooperation Treaty Application No. PCT/US91/07952, the disclosure of which is inco ⁇ orated herein by reference.
  • Suitable media including culture media, lumen media, shell media, and cell perfusion media can be prepared and sterilized using known techniques. As described previously, the recovered cells can be combined with type I collagen in buffer and the resulting solution used to charge (i.e. , load) one or more bioreactors.
  • Culture circuit 14 is operated in the course of media circulation for conditioning the cells in the bioreactor prior to the hemoperfusion procedure, when the patient's blood is circulated through the bioreactor.
  • a bellows pump 28 pulls media from reservoir 30 and circulates it through the switching valve, GEX, pH probe, first and second bioreactors, and a PO 2 probe 32, before returning to the reservoir.
  • the shell media is circulated through the bioreactors, e.g., at a flow of about 500 ml/min, for a period of about 20 to about 24 hours. Over this time the cells contract and the collagen gel forms and constricts, thereby opening up the lumen of the hollow fibers. After completion of the conditioning period, lumen media flow can be established, and the system used for hemoperfusion.
  • a separate lumen-in/lumen-out fluid path 40 provides a dedicated fluid pump 42 for each bioreactor lumen-in and a dedicated fluid pump 44 for each bioreactor lumen-out. This insures equal fluid volume to be pumped in and out of the bioreactors with little fluid loss or gain across the membrane. The effect of this arrangement is to prevent the semipermeable membrane from acting as an ultrafiltration device and consequently to restrict membrane transport of solutes to diffusion controlled processes.
  • Hemoperfusion circuit 16 is operated by placing the switching valve in the appropriate "Patient” or “Hemoperfusion” mode, in order to terminate the flow of media and begin the flow of blood through the circuit.
  • the flow circuit Prior to beginning the hemoperfusion mode, the flow circuit is optionally, and preferably, primed or cleaned, e.g., by vacuum and/or flushing with the patient's blood or another suitable material.
  • a blood pump (not shown) is used to circulate the patient's blood, using standard dialysis blood line 36 through the switching valve, GEX, pH probe, first and second bioreactors, and finally back to the patient via a standard dialysis venous blood line 38.
  • Blood is allowed to flow through the extraluminal shell space, with the semi-permeable hollow fibers serving to separate the blood from the hepatocytes.
  • An intraluminal stream of medium is allowed to flow through the hollow fibers, to carry the growth factors and nutrients necessary to support cell growth and to provide toxin or metabolic product removal.
  • the selectively permeable hollow fibers allow diffusion of toxic waste products, such as ammonia and bilirubin from the blood to the intraluminal biomatrix.
  • the blood composition including the pH and/or blood gas levels can be continually monitored in order to ensure that they are within suitable limits. Variations of these parameters can be achieved by appropriate control of the gas composition and/or flow rate in the oxygenator.
  • the three- dimensional extracellular matrix allows high density cell inoculation and promotes differentiated metabolic function and viability.
  • the hollow fiber membrane molecular weight cut off of 100 kDA provides immunoprotection for device borne xenohepatocytes during whole blood perfusion. Bioartificial liver hemoperfusion using such a system appears to provide a source of differentiated hepatic function, prevent the onset of coma, and improve survival.

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Abstract

Cette invention se rapporte à un ensemble de matériel de culture jetable (10) (c'est-à-dire un module de support de bio-réacteurs (11) et un jeu de tubes (12)) cet ensemble étant conçu pour être utilisé avec un foie bio-artificiel utilisant un bio-réacteur à fibres creuses (24 et 26) comportant un canal d'écoulement extra-capillaire relié à la fois à un circuit (milieu) de culture (14) et à un circuit (sanguin) d'hémoperfusion (16). Cet ensemble (10) comprend une cartouche d'échange gazeux (20) reliée de façon opérationnelle à la fois au circuit de culture (14) et au circuit d'hémoperfusion (16), pour réguler l'acheminement de gaz dans le milieu de culture et dans le sang avant la libération de ceux-ci dans le bio-réacteur (24 et 26). L'utilisation d'une seule cartouche d'échanges gazeux (20) à la fois dans le circuit du milieu de culture (14) et dans le circuit sanguin (16) présente des avantages en termes à la fois de fonctionnalité et de performances par rapport aux autres approches.
PCT/US1996/017559 1995-10-30 1996-10-30 Materiel de culture pour foie bio-artificiel WO1997016527A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP9517579A JPH10512159A (ja) 1995-10-30 1996-10-30 生物人工肝臓のための培養製品
EP96938724A EP0801674A4 (fr) 1995-10-30 1996-10-30 Materiel de culture pour foie bio-artificiel

Applications Claiming Priority (2)

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US55016295A 1995-10-30 1995-10-30
US08/550,162 1995-10-30

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Cited By (20)

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US5955353A (en) * 1997-05-22 1999-09-21 Excorp Medical, Inc. Hollow fiber bioreactor with an extrafilament flow plug
EP1045024A1 (fr) * 1999-04-12 2000-10-18 K.U. Leuven Research & Development Utilisation d'une lignée céllulaire d' hépatocytes humaines pour le traitement de la défaillance hépatique aigue et chronique
WO2003085101A1 (fr) * 2002-04-01 2003-10-16 Cannon Thomas F Systeme automatise pour la bioculture et pour realiser des experiences dans le domaine de la bioculture
CN100335142C (zh) * 2004-12-24 2007-09-05 浙江大学 适用于人工肝的装置
US7270996B2 (en) 2000-10-02 2007-09-18 Cannon Thomas F Automated bioculture and bioculture experiments system
US8309347B2 (en) 2007-03-05 2012-11-13 Terumo Bct, Inc. Cell expansion system and methods of use
US9057045B2 (en) 2009-12-29 2015-06-16 Terumo Bct, Inc. Method of loading and distributing cells in a bioreactor of a cell expansion system
US9617506B2 (en) 2013-11-16 2017-04-11 Terumo Bct, Inc. Expanding cells in a bioreactor
US9677042B2 (en) 2010-10-08 2017-06-13 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US20180044624A1 (en) * 2015-02-20 2018-02-15 Tokyo Electron Limited Cell Culture Device, Cartridge for Culture Medium Replacement Use, and Method for Replacing Culture Medium
US10077421B2 (en) 2014-04-24 2018-09-18 Terumo Bct, Inc. Measuring flow rate
US11008547B2 (en) 2014-03-25 2021-05-18 Terumo Bct, Inc. Passive replacement of media
US11104874B2 (en) 2016-06-07 2021-08-31 Terumo Bct, Inc. Coating a bioreactor
US11312935B2 (en) 2017-01-20 2022-04-26 Nihon Kohden Corporation Cell culture system, cell culture environment evaluation device, and program
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion

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WO1992007615A1 (fr) 1990-10-29 1992-05-14 Regents Of The University Of Minnesota Foie bio-artificiel
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US4889812A (en) * 1986-05-12 1989-12-26 C. D. Medical, Inc. Bioreactor apparatus
WO1992007615A1 (fr) 1990-10-29 1992-05-14 Regents Of The University Of Minnesota Foie bio-artificiel
US5270192A (en) * 1991-02-07 1993-12-14 Monsanto Company Biological artificial liver
US5368555A (en) * 1992-12-29 1994-11-29 Hepatix, Inc. Organ support system

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Cited By (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5955353A (en) * 1997-05-22 1999-09-21 Excorp Medical, Inc. Hollow fiber bioreactor with an extrafilament flow plug
EP1045024A1 (fr) * 1999-04-12 2000-10-18 K.U. Leuven Research & Development Utilisation d'une lignée céllulaire d' hépatocytes humaines pour le traitement de la défaillance hépatique aigue et chronique
US7270996B2 (en) 2000-10-02 2007-09-18 Cannon Thomas F Automated bioculture and bioculture experiments system
US7906323B2 (en) 2000-10-02 2011-03-15 Tissue Genesis, Inc. Automated bioculture and bioculture experiments system
WO2003085101A1 (fr) * 2002-04-01 2003-10-16 Cannon Thomas F Systeme automatise pour la bioculture et pour realiser des experiences dans le domaine de la bioculture
CN100335142C (zh) * 2004-12-24 2007-09-05 浙江大学 适用于人工肝的装置
US8309347B2 (en) 2007-03-05 2012-11-13 Terumo Bct, Inc. Cell expansion system and methods of use
US8785181B2 (en) 2007-03-05 2014-07-22 Terumo Bct, Inc. Cell expansion system and methods of use
US9260698B2 (en) 2007-03-05 2016-02-16 Terumo Bct, Inc. Cell expansion system and methods of use
US9057045B2 (en) 2009-12-29 2015-06-16 Terumo Bct, Inc. Method of loading and distributing cells in a bioreactor of a cell expansion system
US10870827B2 (en) 2010-10-08 2020-12-22 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11613727B2 (en) 2010-10-08 2023-03-28 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US9725689B2 (en) 2010-10-08 2017-08-08 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11773363B2 (en) 2010-10-08 2023-10-03 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11746319B2 (en) 2010-10-08 2023-09-05 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US9677042B2 (en) 2010-10-08 2017-06-13 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US10669519B2 (en) 2010-10-08 2020-06-02 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11667876B2 (en) 2013-11-16 2023-06-06 Terumo Bct, Inc. Expanding cells in a bioreactor
US9617506B2 (en) 2013-11-16 2017-04-11 Terumo Bct, Inc. Expanding cells in a bioreactor
US10633625B2 (en) 2013-11-16 2020-04-28 Terumo Bct, Inc. Expanding cells in a bioreactor
US11708554B2 (en) 2013-11-16 2023-07-25 Terumo Bct, Inc. Expanding cells in a bioreactor
US10557112B2 (en) 2013-11-16 2020-02-11 Terumo Bct, Inc. Expanding cells in a bioreactor
US11008547B2 (en) 2014-03-25 2021-05-18 Terumo Bct, Inc. Passive replacement of media
US11795432B2 (en) 2014-03-25 2023-10-24 Terumo Bct, Inc. Passive replacement of media
US10077421B2 (en) 2014-04-24 2018-09-18 Terumo Bct, Inc. Measuring flow rate
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US20180044624A1 (en) * 2015-02-20 2018-02-15 Tokyo Electron Limited Cell Culture Device, Cartridge for Culture Medium Replacement Use, and Method for Replacing Culture Medium
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion
US11634677B2 (en) 2016-06-07 2023-04-25 Terumo Bct, Inc. Coating a bioreactor in a cell expansion system
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11104874B2 (en) 2016-06-07 2021-08-31 Terumo Bct, Inc. Coating a bioreactor
US11999929B2 (en) 2016-06-07 2024-06-04 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11312935B2 (en) 2017-01-20 2022-04-26 Nihon Kohden Corporation Cell culture system, cell culture environment evaluation device, and program
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
US11702634B2 (en) 2017-03-31 2023-07-18 Terumo Bct, Inc. Expanding cells in a bioreactor

Also Published As

Publication number Publication date
EP0801674A4 (fr) 1999-07-07
EP0801674A1 (fr) 1997-10-22
JPH10512159A (ja) 1998-11-24

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