WO2015160952A1 - Appareil ou dispositif pour thérapie cellulaire - Google Patents

Appareil ou dispositif pour thérapie cellulaire Download PDF

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Publication number
WO2015160952A1
WO2015160952A1 PCT/US2015/025973 US2015025973W WO2015160952A1 WO 2015160952 A1 WO2015160952 A1 WO 2015160952A1 US 2015025973 W US2015025973 W US 2015025973W WO 2015160952 A1 WO2015160952 A1 WO 2015160952A1
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Prior art keywords
cells
disease
tissue
subject
organ
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PCT/US2015/025973
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English (en)
Inventor
Mark Frings PITTENGER
Pablo Gerardo SANCHEZ
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Pittenger Mark Frings
Sanchez Pablo Gerardo
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Publication of WO2015160952A1 publication Critical patent/WO2015160952A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/04—Macromolecular materials
    • A61L29/041—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
    • A61L29/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/64—Animal cells

Definitions

  • hMSCs human mesenchymal stem cells
  • GvHD graft vs host disease
  • MSCs have been directly injected into organs (heart, muscle, kidney), joints for osteoarthritis, and infused systemically for support of hematopoietic transplant, therapeutic treatment of autoimmune diseases, GvHD, and to modify the course of disease after organ damage such as cardiac infarction, chronic heart disease, and acute respiratory distress.
  • the hMSCs produce many soluble bioactive factors that may be responsible for their broad application (Haynesworth et al., J. Cell. Physiol. 166:585-92, 1996; Majumdar et al., J. Cell. Phys. 176:57- 66, 1998; Pittenger et al., Science 284:143-147, 1999; Aggarwal and Pittenger, Blood 105:1815-1822, 2005; Klyushenkova et al., J. Biomedical Sciences 12:47-57, 2005; Di Nicola et al., Blood 99:3838-43, 2002).
  • hMSCs Because of the many therapeutic uses for hMSCs, there is strong interest in understanding the therapeutic capabilities of hMSCs and finding standardized methods for their safe and effective use. However, due to differences in the source of hMSCs and lab procedures, there is limited agreement on the identity, purity, potency, or recommended dose of such cells. Furthermore, delivery of hMSCs by systemic infusion or needle injection to solid organs results in loss of control and absence of knowledge of where the cells are or what they are doing in the body after infusion. Also, there is no method to remove the delivered cells from the body (other than transduction with a suicide gene).
  • the invention features a cell therapy apparatus or device containing an immortalized cell (e.g., immortalized stem or progenitor cells) that can be connected to the blood circulation system of an organ, tissue, or recipient.
  • the immortalized stem or progenitor cells produce therapeutic substances, such as cytokines, chemokines, anti-inflammatory agents, and exosomes, that can be infused to repair a damaged or deficient organ, tissue, or cell (e.g., lung, heart, kidney, liver, and other organs, tissue, or cells).
  • the cell therapy apparatus or device can be used to treat focused tissue damage of an organ, tissue, or recipient, or diffused disease or tissue damage such as sepsis or graft vs host disease.
  • the cell therapy apparatus or device retains the immortalized stem or progenitor cells and only transfers the therapeutic substances produced by the immortalized stem or progenitor cells to the organs, tissues, or cells.
  • the apparatus or device contains a porous material (e.g., a matrix) on which the cells reside.
  • the volume of the apparatus or device is such that it can contain a therapeutically useful number of cells (e.g., in the range of about 50,000 to about 200 million (e.g., about 500,000 to about 200 million)).
  • the apparatus or device may contain any kind of cell with useful properties; particularly useful are
  • immortalized cells such as immortalized stem or progenitor cells.
  • the invention features an apparatus or device containing an inlet port and an outlet port for connection to an organ, tissue, or subject, in which the apparatus or device further contains immortalized cells resident on a matrix in the apparatus or device.
  • the apparatus or device is adapted for connection to the blood circulation system of an organ, tissue, or recipient.
  • the apparatus or device includes two or more inlet ports or two or more outlet ports.
  • the apparatus or device is for ex vivo use or implanted in the subject.
  • the immortalized cells in the apparatus or device are stem or progenitor cells.
  • the cells in the apparatus or device produce effluent medium, wherein said effluent medium is conditioned by the cells.
  • the immortalized cells in the apparatus or device are cells of the lung, heart, liver, bladder, brain, nervous system tissue, blood vessels, skin, eye structures, gut, bone, muscle, ligament, cartilage, esophagus, pancreas, intestines, gallbladder, bile duct, fallopian tubes, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureter, urethra, uterus, or fat.
  • the stem or progenitor cells are embryonic stem cells, induced pluripotent stem cells (IPS), hematopoietic stem cells, intestinal stem cells, osteoblastic stem cells, mesenchymal stem cells (MSCs), multipotent adult progenitor cells (MAPCs), neural stem cells, epithelial stem cells, bone stem cells, cardiac myocyte progenitor stem cells, skin stem cells, skeletal stem cells, muscle stem cells, endothelial stem cells, endothelial progenitor cells, umbilical cord stem cells, adipose stem cells, placental stem cells, placental-derived multipotent stem cells, or liver stem cells.
  • IPS induced pluripotent stem cells
  • hematopoietic stem cells hematopoietic stem cells
  • intestinal stem cells intestinal stem cells
  • osteoblastic stem cells mesenchymal stem cells (MSCs), multipotent adult progenitor cells (MAPCs)
  • neural stem cells epithelial stem cells
  • the mesenchymal stem cells are selected from the group consisting of lung mesenchymal stem cells, Wharton's Jelly mesenchymal stem cells, bone marrow mesenchymal cells, bone marrow stromal cells, umbilical cord mesenchymal cells, spleen mesenchymal cells (e.g., Hox1 1 + , CD45 " cells), adipose derived mesenchymal cells, and pericytes.
  • the epithelial stem cells are selected from the group consisting of lung epithelial stem cells, breast epithelial stem cells, vascular epithelial stem cells, and intestinal epithelial stem cells.
  • the skin stem cells are selected from the group consisting of epidermal stem cells, follicular stem cells, and follicle bulge stem cells.
  • the neural cells are selected from the group consisting of neuronal dopaminergic stem cells and motor neuron stem cells.
  • the stem cells in the apparatus or device are mesenchymal stem cells (MSCs).
  • MSCs mesenchymal stem cells
  • the inlet port or outlet port of the apparatus or device has one or more of a pre-filter, intermediate filter, or final filter.
  • one or more of the pre-filter, intermediate filter, and final filter of the inlet port or outlet port has a filter size in the range of 0.2 - 1 ⁇ .
  • the pre-filter, intermediate filter, and final filter have a filter size of 1 ⁇ , 0.48 ⁇ , and 0.22 ⁇ , respectively.
  • the matrix of the apparatus or device includes polyvinyl acetal, polylactic acid (PLA), polyglycolic acid (PGA), poly(lactic/glycolic acid) (PGLA), hollow fiber substrate, titanium, MATRIGEL® (e.g., reconstituted basement membrane extract that is rich in one or more of laminin, growth factors, entactin/nidogen, type I, II, III, IV, or V collagen, and heparan sulfate proteoglycan (perlecan)), fibronectin, or gelatin.
  • PLA polylactic acid
  • PGA polyglycolic acid
  • PGLA poly(lactic/glycolic acid)
  • hollow fiber substrate titanium
  • MATRIGEL® e.g., reconstituted basement membrane extract that is rich in one or more of laminin, growth factors, entactin/nidogen, type I, II, III, IV, or V collagen, and heparan sulfate proteoglycan (perlecan)
  • the polyvinyl acetal, polylactic acid, or polyglycolic acid is a foam or woven or nonwoven fabric material.
  • the apparatus or device is constructed from polycarbonate, polyacrylate, polystyrene, polysulfone, polyester, poly(methyl methacrylate) (PMMA), polymethacrylate (PMA), poly tetrafluoroethylene (PTFE), or fluorinated ethylene propylene copolymer (FEP).
  • PMMA poly(methyl methacrylate)
  • PMA polymethacrylate
  • PTFE poly tetrafluoroethylene
  • FEP fluorinated ethylene propylene copolymer
  • the apparatus or device is constructed from polycarbonate or fluorinated ethylene propylene copolymer (FEP).
  • FEP fluorinated ethylene propylene copolymer
  • the apparatus or device is constructed of flat sheets of material and fused along the edges to create a bag.
  • the bag contains a matrix.
  • the apparatus or device includes a straight, U-shaped, Z-shaped, or multichannel fluid flow path.
  • the apparatus or device utilizes the Z-shaped fluid flow path.
  • the apparatus or device has a length of between 1 to 5 inches (e.g., 2 to 5 inches), a width of between 1 to 10 inches (e.g., 3 to 10 inches), and a thickness of between 0.2 to 1 .5 inches; particularly the apparatus or device is approximately 3 inches by 5 inches by 0.75 inches.
  • the apparatus or device includes a volume of approximately 1 to 100 cm 3
  • the apparatus or device is constructed of a hard or resilient material. In alternative embodiments, the apparatus or device is constructed of a soft or flexible material. In other embodiments, the apparatus or device includes a tube (e.g., a catheter) connected to the one or more inlet ports or the one or more outlet ports.
  • the tube can be made from, e.g., latex, silicone,
  • the catheter is a Foley catheter, straight catheter, or Quinton catheter.
  • the tube may also include an adapter for attaching the tube to other devices.
  • the apparatus or device further includes an appropriate cell culture medium for the cells.
  • the cell culture medium is selected from phosphate-buffered saline (PBS), Dulbecco's Modified Eagle Medium (DMEM), F-12 medium, a Modified Eagles Medium (a- MEM), or any one or a mixture of these media supplemented with fetal bovine serum (FBS) or human serum, blood, or one or more blood products such as platelet-rich plasma, serum albumin, or recombinant serum albumin.
  • PBS phosphate-buffered saline
  • DMEM Dulbecco's Modified Eagle Medium
  • F-12 medium F-12 medium
  • a- MEM Modified Eagles Medium
  • the cell culture medium in the apparatus or device is PBS.
  • the apparatus or device further includes a heating unit that maintains the immortalized cells in the apparatus or device at a temperature in the range of about 20-37°C (e.g., 30- 37°C).
  • the apparatus or device is connected to an organ or tissue of a subject or to the circulatory system of the subject.
  • the organ or tissue is in vivo or ex vivo.
  • the apparatus or device is connected to the circulatory system of the subject.
  • the apparatus or device is connected extracorporeally to the subject.
  • the apparatus or device is implanted into the subject.
  • the apparatus or device is connected immediately proximal to the organ or tissue.
  • the subject is a transplant recipient, and the apparatus or device is connected to the subject in preparation for transplantation of an organ or tissue.
  • the organ or tissue connected to the apparatus or device is being prepared for transplantation into a recipient.
  • the apparatus or device including immortalized cells is infused with a bioactive factor or a cell that causes the immortalized cells (e.g., the resident, immortalized cells) in the apparatus or device to produce one or more molecules not otherwise made in the apparatus or device.
  • the bioactive factor is TGFfi or TGFa.
  • the cell that causes the immortalized cells in the apparatus or device to produce one or more molecules not otherwise made in the apparatus or device is a peripheral blood mononuclear cell
  • the immortalized cells in the apparatus or device are bioengineered to produce one or more molecules. In some embodiments, the immortalized cells in the apparatus or device are bioengineered to produce insulin.
  • the apparatus or device includes a biological sample from a donor or a recipient.
  • the biological sample may include one or more cells, fluid, or bioactive factors obtained from the donor or the recipient.
  • the biological sample is injected into the apparatus or device.
  • the organ or tissue connected to the apparatus or device is from lung, heart, liver, bladder, brain, blood vessels, skin, eye structures, gut, bone, muscle, ligament, cartilage, esophagus, pancreas, intestines, gallbladder, bile duct, fallopian tubes, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureter, urethra, or uterus.
  • the apparatus or device is connected to a container that includes stem or progenitor cells isolated from a subject.
  • the subject or recipient is a human or a non-human mammal. Particularly, the subject or recipient is a human.
  • the organ or tissue to which the apparatus or device is connected is a human organ or tissue.
  • the apparatus or device promotes proliferation of the stem or progenitor cells.
  • the apparatus or device can be used to promote proliferation of stem or progenitor cells, such as hematopoietic stem cells, inside the device, or outside the device (e.g., in vitro or in vivo).
  • the immortalized cells in the apparatus or device are human cells, particularly the human cells are human stem or progenitor cells, such as human MSCs (e.g., human bone marrow MSCs).
  • human MSCs e.g., human bone marrow MSCs
  • a second aspect of the invention features a method of treating a medical condition by establishing a fluid communication between the apparatus or device of the first aspect of the invention and an organ, tissue or subject.
  • the subject is a human or a non-human mammal; particularly, the subject is a human.
  • the organ or tissue is a human organ or tissue.
  • the method uses immortalized cells, such as human cells; particularly the human cells are human stem or progenitor cells, such as human MSCs (e.g., human bone marrow MSCs).
  • human MSCs e.g., human bone marrow MSCs
  • the organ or tissue is in a subject or is ex vivo; particularly the organ or tissue is in the subject.
  • a third aspect of the invention features a method of treating a medical condition in a subject including collecting the effluent medium produced by the cells in the apparatus or device of the invention and administering the effluent medium to the subject, wherein the effluent medium is conditioned by the cells.
  • a fourth aspect of the invention features a method for analyzing and identifying for molecules in the effluent medium produced by the cells in the apparatus or device of in the invention including collecting the effluent medium, detecting in the effluent medium one or more molecules produced by the cells, and identifying and isolating the one or more molecules produced by the cells, wherein the effluent medium is conditioned by the cells.
  • the methods further include infusing the apparatus or device of the invention with a bioactive factor or a cell that causes the immortalized cells in the apparatus or device to produce one or more molecules not otherwise made in the apparatus or device.
  • the bioactive factor is TGFfi or TGFa.
  • the cell that causes the immortalized cells in the apparatus or device to produce one or more molecules not otherwise made in the apparatus or device is a peripheral blood mononuclear cell
  • the method is used to treat a medical condition.
  • the medical condition is an acute disease, a chronic disease, an autoimmune disease, a skin disease, a neurodegenerative disease, a musculoskeletal disease, a muscle wasting disease, cancer, a vascular or circulatory disease, an inflammatory condition, a cytokine storm condition, an immunological condition, graft versus host disease, a rheumatology disease, cardiac infarction, acute or chronic heart disease, acute or chronic respiratory disease or distress, pulmonary fibrosis, trauma, sepsis, or organ failure.
  • the autoimmune disease is Alopecia Areata, Ankylosing Spondylitis,
  • Antiphospholipid Syndrome Addison's Disease, Hemolytic Anemia, Hepatitis, Behcets Disease, Bullous Pemphigoid, Cardiomyopathy, Celiac Sprue-Dermatitis, Chronic Fatigue Immune Dysfunction Syndrome (CFIDS), Chronic Inflammatory Demyelinating Polyneuropathy, Churg-Strauss Syndrome, Cicatricial Pemphigoid, Limited Scleroderma (CREST Syndrome), Cold Agglutinin Disease, Crohn's Disease, Discoid Lupus, Essential Mixed Cryoglobulinemia, Fibromyalgia-Fibromyositis, Graves' Disease, Guillain- Barre Syndrome, Hashimoto's Thyroiditis, Hypothyroidism, Idiopathic Pulmonary Fibrosis, Idiopathic
  • IgA Thrombocytopenia Purpura
  • IgA IgA Nephropathy, insulin dependent diabetes, Juvenile Arthritis, Lichen Planus, Lupus, Meniere's Disease, Mixed Connective Tissue Disease, Multiple Sclerosis, Myasthenia Gravis, Pemphigus Vulgaris, Pernicious Anemia, Polyarteritis Nodosa, Polychondritis, Polyglandular Syndromes, Polymyalgia Rheumatica, Polymyositis and Dermatomyositis, Primary Agammaglobulinemia, Primary Biliary Cirrhosis, Psoriasis, Raynaud's Phenomenon, Reiter's Syndrome, Rheumatic Fever, Rheumatoid Arthritis, Sarcoidosis, Scleroderma, Sjogren's Syndrome, Stiff-Man Syndrome, Takayasu Arteritis, Temporal Arteritis/Giant Cell Arteritis, Ulcerative Co
  • the neurodegenerative disease is a neurological disorder selected from the group consisting of Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, cerebral stroke, traumatic brain injury, and spinal cord injury.
  • the neurological disease may be the result of an autoimmune condition.
  • the cancer is bladder cancer, pancreatic cancer, cervical cancer, lung cancer, liver cancer, ovarian cancer, colon cancer, stomach cancer, virally induced cancer,
  • neuroblastoma a tumor necrosis originating from breast cancer, prostate cancer, renal cancer, leukemia, sarcomas, myeloma, or a carcinoma.
  • the method treats cachexia, sarcopenia, reduces chemotherapy-induced inflammation, reduces sepsis, or reduces cancer cell proliferation.
  • the inflammatory condition is osteoarthritis, transplantation related inflammation, cytokine storm, ulcerative colitis, Crohn's disease, proctitis, microscopic colitis, allergic eosinophilic gastroenteritis, food allergies, pill induced esophagitis, celiac disease, recurrent polyps, hemorrhoids, bacterial sepsis, sterile inflammation, an immunological condition, graft versus host disease, or trauma related tissue injury.
  • the method prepares the organ or tissue for transplantation or improves transplantation outcome.
  • the organ or tissue is in vivo or ex vivo.
  • the organ is or is from lung, heart, liver, bladder, brain, blood vessels, skin, eye structures, gut, bone, muscle, ligament, cartilage, esophagus, pancreas, intestines, gallbladder, bile duct, fallopian tubes, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureter, urethra, or uterus.
  • the method promotes the proliferation of cells in the organ, tissue, or subject, such as hematopoietic stem cells.
  • a fifth aspect of the invention features a kit including the apparatus or device of the first aspect of the invention and a cell culture medium or PBS.
  • the cell culture medium is in the apparatus or device.
  • the cell culture medium is in a container in the kit.
  • the kit further comprises a heating unit.
  • the immortalized cells are present in the apparatus or device or are present in a container within the kit.
  • the kit further comprises a reservoir for collecting effluent medium.
  • the effluent medium can be used for analysis, research, or therapeutic use.
  • subject means a vertebrate, such as a mammal, such as a human. Mammals include, but are not limited to, humans, dogs, cats, horses, cows, and pigs.
  • matrix material includes any material useful as a cell support (e.g., in a device or apparatus of the invention) that allows the flow of PBS or medium around the attached cells.
  • hollow fiber is intended to include hollow structures (of any shape) containing pores of defined size, shape and density for use in delivering nutrients (in solution) to cells contained within an apparatus or device of the invention and for allowing bioactive molecules (in solution) produced from cells contained within the apparatus or device to flow out of the apparatus or device and into an organ, tissue, or subject connected to the apparatus or device.
  • hollow fibers may be constructed of a resorbable or nonresorbable material. Fibers include, but are not limited to, tubular structures.
  • induced pluripotent stem cells means somatic cells that have been
  • IPS cells have been derived using modifications of an approach originally discovered in 2006 (Takahashi et al., Cell 131 :861 -872, 2007; Yamanaka, S. et al., Cell Stem Cell, 1 :39-49, 2007). For example, in one instance, to create IPS cells, scientists started with skin cells that were then modified by a standard laboratory technique using retroviruses to insert genes into the cellular DNA.
  • the inserted genes were Oct4, Sox2, Klf4, and c-myc, known to act together as natural regulators to keep cells in an embryonic stem cell-like state. These cells have been described in the literature. See, for example, Wernig et al., PNAS, 105:5856-5861 (2008); Jaenisch et al., Cell, 132:567-582 (2008); Hanna et al., Cell, 133:250-264 (2008); and Brambrink et al., Cell Stem Cell, 2:151 -159 (2008).
  • iPS cells can be created after mRNA for the reprogramming factors are introduced into cells (Warren et al., Cell Stem Cell 7:618-630, 2010).
  • FIG. 1 describes several cell therapy considerations regarding traditional cell therapy using exogenous cell infusion (left) and cell therapy of the invention using an ex vivo cell therapy apparatus or device of the invention (right).
  • Cell therapy considerations 2 to 10 associated with traditional cell therapy may be collapsed into a single consideration when the cell therapy apparatus or device is used, which saves time and decreases the complexity of cell therapy.
  • FIG. 2A is an image showing a prototype cell therapy apparatus or device (e.g., which can be used ex vivo) that can hold 50-150 million MSCs.
  • An inlet port is shown at the upper left and an outlet port is shown at the lower right.
  • the white material is a matrix for cell attachment.
  • the direction of fluid flow through the apparatus or device is depicted by the arrows.
  • the outlet port also includes two tandem filters to prevent any cells from the device from transferring to the tissue, organ, or recipient.
  • FIG. 2B is an image of another prototype cell therapy apparatus or device (e.g., which can be used ex vivo) constructed of sheets of fluorinated ethylene propylene copolymer (FEP).
  • FEP fluorinated ethylene propylene copolymer
  • FIGs. 3A and 3B are images showing that matrix attachment improves the viability of MSCs in vivo.
  • FIG. 3A shows allogeneic MSCs infused by catheter in a dog infarct model and imaged in vivo. Only a small portion of the MSCs homed to the infarct and the detectable number of MSCs decreased by 99% in one week (Kraitchman et al., Arterioscler. Thromb. Vase. Biol. 29:1025-1030, 2009).
  • FIG. 3B shows that allogeneic MSCs persist at 16 weeks when implanted on a matrix. Shown are 100s of CM-Di-I labeled allogeneic MSCs on HA/TCP matrix.
  • FIG. 5 is an illustration showing the factors from hMSCs that modulate immune responses and bacterial sepsis.
  • FIG. 6 is a photograph showing the size of a therapeutic dose of 100 million hMSCs.
  • FIG. 7 is a schematic showing an ex vivo cell therapy system within the scope of the invention.
  • the schematic shows a reservoir, (R), containing PBS or nutrient media which flows into the apparatus or device from the top.
  • Cell stimulating molecules and/or drugs can be added to the fluid contained in the reservoir.
  • the apparatus or device provides (A): a porous matrix seeded with immortalized cells, which continuously produce important molecules into the media or can be stimulated to make additional molecules.
  • the apparatus or device retains the immortalized cells as resident cells and maintains their viability.
  • the apparatus or device also provides (B): a triple filter in the outlet port containing a 1 ⁇ pre- filter, a 0.48 ⁇ intermediate filter, and a 0.22 ⁇ final filter.
  • the triple filter allows the therapeutic molecules produced by the immortalized cells in the apparatus or device to be transferred to the subject.
  • the transferring process can be done using, e.g., a fluid pump (C).
  • C a fluid pump
  • the schematic also shows that the ex vivo cell therapy system can be attached in many ways to a subject (D), such as to the subject's circulation by way of a central vein or a peripheral vein or to the subject's organ or tissue by way of a catheter.
  • the invention features an apparatus or device for cell therapy.
  • the apparatus or device contains immortalized cells (e.g., immortalized stem or progenitor cells, such as human stem or progenitor cells, in particular, human MSCs) on a matrix that can be used to deliver bioactive factors produced by the immortalized cells into a subject, organ, or tissue when the apparatus or device is fluidly connected to the subject, organ, or tissue (e.g., via an inlet port and an outlet port, which can be connected to a major or minor artery or vein, or to other vessels, or to a collecting sinus of the organ or tissue).
  • the apparatus or device may be implanted or present outside the body of a subject (extracorporeal) and attached to an in vivo organ or tissue via tubing or catheters.
  • the attachment may occur proximal or distal to the in vivo organ or tissue of the subject that needs treatment.
  • the apparatus or device may also be connected to the subject such that fluid from the apparatus or device flows into the subject's systemic circulation.
  • the apparatus or device may also be attached directly to an ex vivo organ or tissue (e.g., an organ or tissue that is removed from a donor, e.g., for transplantation into a recipient).
  • an ex vivo organ or tissue e.g., an organ or tissue that is removed from a donor, e.g., for transplantation into a recipient.
  • the apparatus or device may be used to improve transplant outcomes.
  • This apparatus or device may also be used to treat other medical conditions, including, e.g., inflammatory conditions, autoimmune disease, respiratory conditions, cardiac conditions, chronic heart disease, acute respiratory distress, trauma, organ failure, sepsis, neurodegenerative conditions, and cancer.
  • Fluid may be circulated through the apparatus or device and into the organ, tissue, or subject.
  • the organ or tissue may be separated from the systemic circulation of the subject, but remain in the patient (e.g., the organ or tissue is not removed from the patient).
  • the apparatus or device may be connected to an organ or tissue that has been removed from a subject (e.g., a donor).
  • the apparatus or device may also be connected to an organ, tissue, or subject for a limited period of time (e.g., during a procedure that lasts 5 minutes to up to 2-5 hours or more (e.g., 24-72 hours or more)).
  • the apparatus or device may also be connected to an organ, tissue, or subject and fluid re-circulated as needed.
  • the device may be turned on and off such that fluid circulation can be controlled without removing the device from the subject.
  • the device may be detached and reattached, as needed.
  • the device can be replaced with a fresh device between treatments, as needed.
  • Clinical Indication The physician must first consider the patients general health, medical history, age, and immediate clinical indications.
  • Identity/Cell Type The physician must identify the appropriate cell therapy and be familiar with the various cell choices, their availability, and characteristics of the cells that make them useful as a therapeutic. Different cells can produce different quantities of cytokines and factors, and cells produced in different batches can have different levels of production. The age of the cells is important, as this can affect their production of bioactive cytokines and factors. As there are many cells that may have therapeutic potential, choosing an appropriate cell type can be a complex consideration.
  • the cell therapy apparatus or device containing immortalized cells solves this problem by providing a consistent cell type that remains similar from batch to batch. For different therapeutic indications, it may be optimal to use a particular type of immortalized cell in the apparatus or device, as is discussed below.
  • the mode of delivery to the patient must be determined.
  • the cells may be delivered alone, in a special medium, on microspheres, or implanted on a solid matrix.
  • the cells may be needle injected locally into damaged tissue or delivered systemically.
  • the cell therapy apparatus or device containing resident immortalized cells solves these problems since the cells are contained and only the cells' products are infused to the recipient via an intravenous (i.v.) connection for systemic delivery or via a catheter that directs the cell products to the tissue or organ of interest to improve repair, function, and regeneration.
  • Toxicity - Toxicity is always a consideration with effective, powerful medicines including cellular therapy.
  • the effective dosage is usually balanced against its toxicity. Since a good portion of the delivered cells die when delivered by current therapies, it is difficult to know whether cell death increases or decreases the associated toxicity. Similarly, the level of toxicity may be changed if more cells survive delivery. Once delivered, there is no way to reduce the dose or the potential toxic effects resulting from apoptosis of the delivered cells.
  • the cell therapy apparatus or device containing resident immortalized cells solves this problem by keeping the cells in a favorable environment so their production of cytokines and growth factors is consistent and predictable. If there are signs of toxicity in the recipient, the apparatus or device can be disconnected or removed and, if necessary, replaced with a "fresh" apparatus or device.
  • Biodistribution - TUe location, action, and survival of the delivered cells are important factors to consider in cell therapy. Once delivered, the cells cannot be removed or be moved to another location inside the body.
  • the cell therapy apparatus or device containing immortalized cells solves the biodistribution problem since cells reside in a single known location (i.e., the apparatus or device). The cells in the apparatus or device can be tested periodically to assess their viability. It is also possible to "relocate" the apparatus or device and cells to another site of interest by making a new vascular connection in the desired location. 7. Degradation - The vast majority of delivered cells do not find a site of engraftment and will die (apoptose) and be removed over time by macrophages.
  • the cell therapy apparatus or device containing immortalized cells solves this problem by keeping the cells in a favorable environment inside the apparatus or device, where their viability can be ascertained and their production of bioactive factors can be analyzed over time.
  • the cell therapy apparatus or device containing resident immortalized cells solves this problem because the immortalized cells can be standardized so that they are substantially the same time and time again in devices used for the same therapy. Furthermore, the cells can be stored frozen, and last almost indefinitely, providing substantially the same standardized cells for use in the apparatus or device over time.
  • 1 Clinical Outcome - As with all therapies, clinical outcome is the most important consideration and is dependent on many of the factors discussed above that are outside of the physician's control.
  • the cell therapy apparatus or device containing resident immortalized cells eases many of the considerations mentioned above and therefore would be able to provide more predictable clinical outcomes.
  • the invention features an ex v/Vo/extracorporeal or implanted cellular therapy apparatus or device that avoids infusion of exogenous cells, but allows their products, e.g., secreted cytokines, chemokines, and growth factors, to be transferred to the recipient or recipient's cells, tissues, or organs. Rather than infusing the cells directly, the cellular therapy apparatus or device retains the cells within the apparatus or device (as resident cells attached to a substrate, e.g., a matrix) and maintains their viability therein.
  • the cell therapy apparatus or device by itself allows easy handling of the cellular therapy material, and prevents the recipient from receiving the cells directly, eliminating many of the perceived risks of cellular therapy discussed above.
  • the cell therapy apparatus or device containing resident immortalized cells substantially eliminates variability in many of the cell parameters that may vary from batch to batch, such that the cellular characteristics of the apparatus or device will be substantially the same each time, which facilitates quality control and reproducibility.
  • the apparatus or device may be connected to the recipient patient via a central or peripheral vein or artery, to a collecting sinus, or via a cut down procedure to access the cells, tissues, or organs of interest. Accordingly, the apparatus or device provides direct infusion of bioactive factors that are secreted by the resident cells of the apparatus or device.
  • the cellular therapy apparatus or device can be removed, discontinued, or detached from the recipient or patient at any time, which is not possible when the cells are directly infused to the recipient or patient.
  • immortalized hMSCs, stem or progenitor cells, or other cells may be used as the resident cells in the apparatus or device.
  • Having the cell therapy apparatus or device should provide greater control and deeper understanding of the activities of the cells (e.g., hMSCs) used during treatment and allow industrial optimization. Two prototypes of this ex-wVo/extracorporeal cellular therapy apparatus or device are available (described in detailed below).
  • the prototypes are provided to exemplify the apparatus or device of the invention, but should not be considered as limiting the design or scope of the apparatus or device of the invention in any way. Because the resident cells in the apparatus or device are not freely infused to the recipient, the cell therapy apparatus or device is inherently safer than most current methods for providing cellular therapy.
  • FIGs. 2A and 2B each shows a non-limiting prototype of a cell therapy apparatus or device of the invention.
  • the size and shape of the apparatus or device is similar to a cell phone (e.g., about 3 inches by 5 inches by 0.75 inches).
  • the Plexiglas ® prototype apparatus or device of FIG. 2A is constructed by machining a cavity to hold a suitable matrix material (white in FIG. 2A) to support resident cell attachment and viability.
  • the matrix material is encased in a clear container that protects the internal constituents.
  • the apparatus or device also contains an inlet, outlet, and Z-shaped fluid flow path to allow fluid to flow through the matrix material.
  • the outlet of the apparatus or device has a 1 ⁇ pre-filter, a 0.48 ⁇ intermediate filter, and a 0.22 ⁇ final filter.
  • a second prototype constructed as a "flexible bag" is shown in FIG. 2B and is made from sheets of fluorinated ethylene propylene co-polymer (FEP) fused at the edges and containing a polyvinyl acetal foam matrix upon which the cells reside.
  • FEP fluorinated ethylene propylene co-polymer
  • the culture medium used in the apparatus or device is pre- gassed with 5% C0 2 and exchanged every few days, or can be continuously perfused by a pump.
  • a constant flow rate of 0 to 1 .5 ml/min is easily achieved with a peristaltic or constant flow pump.
  • a programmable fluid flow/fluid warmer is available and can be used to maintain the apparatus or device at a constant temperature (e.g., in the range of, e.g., 20-37°C (e.g., 30-37°C).
  • a constant temperature e.g., in the range of, e.g., 20-37°C (e.g., 30-37°C).
  • FIG. 7 A schematic illustrating an example of an ex vivo cell therapy system within the scope of the invention is shown in FIG. 7.
  • the apparatus or device can be made in different sizes and contain 50,000-200 million, such as 50- 200 million, immortalized cells.
  • a typical therapeutic dose for heart ischemia or lung injury is about 100 million hMSCs.
  • the photograph in FIG. 6 shows the size of a therapeutic dose of 100 million hMSCs.
  • the cell therapy apparatus or device could be available off-the-shelf to provide a safe treatment, which could be started, stopped, and disconnected when treatment is completed, which is not the case for injected cells.
  • the extended viability of the cells in the cell therapy apparatus or device compared to
  • FIGs. 3A and 3B show that matrix attachment improves the viability of MSCs in vivo. Given the large number of cellular therapy clinical trials underway, the cell therapy apparatus or device containing immortalized cells may be an important addition to the arena of cellular therapy.
  • the apparatus or device retains the exogenous immortalized cells (e.g., hMSCs) as resident cells and transfers the therapeutic substances, e.g., bioactive factors, produced by the immortalized cells such as cytokines, chemokines, and anti-inflammatory agents to the cells, tissues, and organs of the recipient.
  • the resident cells can maintain viability on many different kinds of suitable matrix materials.
  • Hollow fiber technology has been available for many years and hMSCs have been grown in such bioreactors. Hollow fiber bioreactors and cells that are grown in such reactors are described in U.S. Patent Publication No. US 20120308531 , which is incorporated by reference in its entirety.
  • the matrix material or hollow fibers for use in the apparatus or device of the invention can be pre- coated with one or more extracellular matrix proteins, for example, MATRIGEL®, laminin, fibronectin, or collagen, to enhance cell attachment.
  • Extracellular matrix proteins may be attached to a cell support (e.g., a matrix material or to the internal and/or external surface of the fibers).
  • extracellular matrix protein may be attached to the surface by any of the methodologies as described in U.S. Pat. No. 5,872,094 and U.S. Pat. No. 6,471 ,689, both of which are incorporated herein by reference for teaching these
  • the matrix material or hollow fibers should be suitable for the delivery of nutrients and removal of biological agents from the apparatus or device to the organ, tissue, or subject to which the apparatus or device is connected.
  • the matrix material or hollow fibers may be any shape, for example, they may be round and tubular or in the form of concentric rings.
  • the matrix material or hollow fibers may be made up of a resorbable or non-resorbable membrane.
  • suitable components of the matrix material or hollow fibers include polydioxanone, polylactide, polyglactin, polyglycolic acid, polylactic acid, PLGA, polyglycolic
  • the criteria for matrix materials includes, but is not limited to, the following: not toxic to the cells; porous for the removal of biological agents and materials and the reception of nutrients and, in instances where collection of components secreted by the cell is desired, porosity is adjusted for that parameter; relatively insensitive to temperature changes, i.e., thermally stable; able to retain shape integrity.
  • MSCs have also been grown on multiple types of matrix materials and scaffolds, even titanium, in vitro and in vivo.
  • Multiple apparatus or device designs may be used for the apparatus or device of the invention (e.g., hollow fiber, non-woven, woven, beads), and many different fluid flow paths (e.g., straight, U- shaped, Z-shaped, and multichannel) may be used.
  • the apparatus or device may be relatively small in size, of simple construction, made from non-proprietary materials, and designed to hold about 50,000-200 million, such as 50-200 million, such as 50-150 million, immortalized cells (e.g., hMSCs).
  • the matrix holding the immortalized cells can be made of several materials, but we have used a nonwoven polylactic acid (PLA), a polyvinyl acetal foam, and a collagen sponge with similar success.
  • the cell therapy apparatus or device may be formed of polycarbonate.
  • the apparatus or device is sealed, and gas sterilized (although other known sterilization techniques may be used).
  • the fluorinated ethylene propylene copolymer (FEP) sealed bag containing the polyvinyl acetal foam is steam sterilized and dried before being loaded with cells.
  • the immortalized cells can be inoculated into the apparatus or device in a biological safety cabinet and the cells may be grown for 7-14 days to achieve a desired number of captured immortalized cells (e.g., im-hMSCs).
  • the immortalized cells e.g., im- hMSCs
  • the immortalized cells can also be grown outside the apparatus or device and then inserted into the apparatus or device once the desired number of cells are ready. It is important to use immortalized cells in the apparatus or device because in order to make a reproducible cell therapy apparatus or device that responds in a consistent fashion, the cells within the apparatus or device should be substantially the same from batch to batch. Newly isolated stem cells can be used each time, but the apparatus or device parameters will likely not be as consistent.
  • the cell therapy apparatus or device connected to the blood supply may need to be just proximal to the organ of interest to offer greatest therapeutic potential. This may require surgical intervention, such as a procedural cut-down to access the needed blood vessel. Surgeons have assured us that this is not difficult and has small but real risks commonly associated with vascular surgery.
  • a cell therapy apparatus or device containing immortalized cells e.g., im- hMSCs
  • the apparatus or device may contain multiple inlet and outlet openings or ports. One or more of the ports may be used to collect effluent medium or add, e.g., materials, fluids, bioactive factors, agents, and cell culture medium to the apparatus or device.
  • the apparatus or device may also be in the form of a culture bag (e.g., a gas-permeable culture bag).
  • the cell therapy apparatus or device as described herein is an early evolution of cellular therapy that does not use direct infusion of therapeutic cells.
  • the described apparatus or device can be used in therapeutic or industrial methods, as well as to improve our current understanding of stem or progenitor cell therapy (e.g., hMSC therapy).
  • the apparatus or device here which may be about the size of a cell phone, may contain up to 200 million (e.g., 5-200 million) immortalized stem or progenitor cells (e.g., hMSCs) that can produce therapeutic effects by transferring their bioactive factors, e.g., cytokines, chemokines, and anti-inflammatory agents, when connected to the patient's circulation or to an organ or tissue.
  • hMSCs immortalized stem or progenitor cells
  • the apparatus or device can be disconnected when treatment is over or restarted as needed without subjecting the patient (or an organ or tissue) to injected cells and their unknown long term effects.
  • somatic stem cells including hematopoietic stem cells and MSCs.
  • Approaches to translate the therapeutic potential of these cells to clinical applications suffer from the lack of supply of suitable cells, their short life-span, and high inter- donor viability within stem/progenitor cell preparations.
  • the low amount of hMSCs in bone marrow extracts typically necessitates an in vitro cell growth phase prior to use. This in vitro cell growth phase is limited by the replicative senescence phenomenon occurring under in vitro culture conditions, after 25 to 60 population doublings (PD).
  • hMSCs differentiation potential
  • the differentiation potential of hMSCs displays significant variations from donor to donor and this may reflect other undesirable variations from preparation to preparation.
  • the low supply of hMSCs combined with their short life-span and the high inter-donor variability in characteristics limits the practical therapeutic potential of these stem cells.
  • Immortalized stem/progenitor cells can be propagated in large numbers with the pluripotency intact, although pluripotency and differentiation potential may not be required for most of the envisioned applications.
  • Immortalized cells e.g., hMSCs
  • hMSCs can be produced using methods known in the art.
  • gene expression levels of immortalized cells e.g., im-hMSCs
  • Immortalized cells or stem cells of the invention are cells that may maintain their multipotencies, produce useful factors, and/or have the capacity to grow indefinitely.
  • Methods of producing immortalized cells include taking non-immortalized cells and introducing an "immortalizing gene" to the genome of the cells.
  • Non-integrating vectors that do not introduce immortalizing genes into chromosomes, such as episomal plasmids and mini-chromosomes, are also known in the field.
  • Methods of producing immortalized cells are well known in the art and are described in detail below or in publications incorporated herein by reference.
  • hMSCs Human mesenchymal stem cells derived from the bone marrow may be used as the resident stem cells in a cell therapy apparatus or device of the invention. These stem cells are defined as flow cytometry positive for CD73, CD90, and CD105, and negative for hematopoietic markers (e.g., CD34) and are able to stably differentiate in vitro into osteoblasts, adipocytes, and chondrocytes (Pittenger et al., Science 284:143-147, 1999).
  • the multidifferentiation capacity of hMSCs makes them promising candidates for regenerative medicine.
  • the chondrogenic and osteogenic potential of hMSCs is useful for the repair of damaged articular cartilage but also for bone tissue engineering by recapitulating intramembranous or endochondral ossification processes.
  • hMSCs can participate in building tissue, particularly bone, they also produce multiple antiinflammatory agents, growth factors, and cytokines, as is described below. Many factors are produced constitutively in substantial amounts by hMSCs, and interaction of hMSCs with other cell types causes additional factor release, either from the hMSCs or the other cells (Haynesworth et al., J. Cell. Physiol. 166:585-92, 1996; Majumdar et al., J. Cell. Phys. 176:57-66, 1998; Aggarwal and Pittenger, Blood 105:1815-1822, 2005; Klyushenkova et al., J.
  • hMSC-derived anti-inflammatory agents have profound effects on the local environment of tissues and organs, but also can have effects systemically, such as in the case of patients suffering from GvHD.
  • hMSCs respond to their neighboring cells and modulate tissue responses gradually and appropriately over time, such as in the case of hMSC modulation of cardiac remodeling when the cells are injected following an infarct (Zhao et al., Stem Cells Transl. Med. 1 :685-695, 2012). It is well-known in the art that hMSCs are biologically responsive to their local environment in vivo. As an example, hMSCs produce vascular endothelial growth factor (VEGF) but upon interaction with peripheral blood mononuclear cells, the amount increases by three fold (Aggarwal and Pittenger, supra).
  • VEGF vascular endothelial growth factor
  • MSC-like cells can be isolated from different tissue sources and produced in different labs and by different commercial organizations, although the cells have similar cell properties, uses, immunological effects, and produce similar bioactive factors.
  • the names given to these MSC-like cells include mesenchymal stromal cells, marrow stromal cells, mesenchymal precursor cells (MPC) (Hamamoto et al., Ann Thorac Surg. 87:794- 801 , 2009), marrow stromal stem cells (Gronthos et al., Journal of Cell Science. 1 16:1827-1835.
  • MPC multipotent adult progenitor cells
  • SkSCs skeletal stem cells
  • stromal stem cells (Gronthos et al., Journal of Cell Science 1 16:1827- 1835. 2003), adipose-derived mesenchymal stem cells, adipose derived stromal stem cells (Zannettino et al., J. Cell. Physiol. 214:413-421 , 2008), multilineage adipose cells (Zuk et al., Tissue Eng. 12:2813-2823, 2001 ), marrow isolated adult multilineage inducible (MIAMI) cells (D'lppolito et al., J. Cell. Sci.
  • MIAMI marrow isolated adult multilineage inducible
  • stem cells from human exfoliated deciduous teeth (SHED) (Miura et al., Proc. Natl. Acad. Sci. U.S.A. 100:5807-5812, 2003; Yamaza et al., Stem Cell Res. Ther. 1 :5, 2010), dental pulp stem cells (DPSCs) (Zuk et al., Tissue Eng. 12:2813-2823, 2001 ), periodontal ligament stem cells (PDLSCs), stem cells from apical papilla (SCAP), and dental follicle precursor cells (DFPCs).
  • SHED human exfoliated deciduous teeth
  • DPSCs dental pulp stem cells
  • PDLSCs periodontal ligament stem cells
  • SCAP apical papilla
  • DFPCs dental follicle precursor cells
  • APCs adult progenitor cells
  • PDAC ® pluripotent placental derived adherent cells
  • PDA-001 or PDA-002 Celgene Cellular Therapeutics, Inc.
  • Celgene Cellular Therapeutics, Inc. culture-expanded MSCs from bone marrow
  • PROCHYMAL ® and CHONDROGEN ® Osiris Therapeutics, Inc.
  • MULTISTEM ® Athersys Inc.
  • placental expanded (PLX) cells Pluristem Therapeutics, Inc.
  • ALD-201 ALD-301 , and ALD-401
  • ALD-401 Cytomedix, Inc.
  • the MSC or MSC-like cells are also reported to stimulate blood vessel formation and the appearance of other important reparative cells such as c-Kit+ stem cells in damaged tissue, most notably the heart (Orlic et al., Proc. Natl. Acad. Sci. U.S.A. 98:10344-10349, 2001 ; Beltrami et al., Cell 1 14:763- 776, 2003).
  • Other stem/progenitor cell populations that have been isolated from hearts and characterized are termed the cardiosphere derived cells (CDCs) and the related cardiosphere progenitor cells (CSpc) (Chimenti et al., Circ. Res. 106:971 -980, 2010).
  • VEGF vascular endothelial growth factor-1
  • HGF HGF
  • IGF-1 insulin-like growth factor-1
  • Hox 1 1 + cells e.g., Hox1 1 + , CD45 " cells from spleen
  • Faustman et al. Feaustman et al., Int. J. Biochem. Cell Biol. 42:1576-1579, 2010; Lonyai et al., Horm. Metab. Res. 40:137-146, 2008; U.S. Patent Publication NOs. 8,021 ,693, 7,582,313, and 8,017,392; incorporated herein by reference.
  • These cells also produce important factors.
  • the Hox 1 1 + cells are under intense study for their ability to form pancreatic islet cells useful for treating diabetes.
  • the cell therapy apparatus or device containing immortalized Hox 1 1 + cells can be useful for the study and treatment of diabetes.
  • These Hox1 1 + cells could also be used in the cell therapy apparatus or device, preferably with the cells immortalized, for use in the treatment of diabetes and other autoimmune disease or to promote repair and regeneration of damaged tissue and organs.
  • any cellular use that involves, as part of its mode of action, the expression of cytokines, growth factors, or other molecules, such as those systems that utilize CD34+ cells or their derivatives, alone or in combination with MSC-like cells, such as ixmyelocel from Aastrom Biosciences Inc., could be combined with the apparatus or device of the invention as is or in an immortalized form.
  • stem cells that can also be used in the invention include, but are not limited to, embryonic stem cells, induced pluripotent stem cells (IPS), hematopoietic stem cells, intestinal stem cells, osteoblastic stem cells, mesenchymal stem cells, multipotent adult progenitor cells (MAPCs), unrestricted somatic stem cells (USSCs), neural stem cells, epithelial stem cells, bone stem cells, cardiac myocyte progenitor stem cells, skin stem cells, skeletal stem cells, placental stem cells, placental-derived multipotent stem cells, muscle stem cells, endothelial stem cells, umbilical cord stem cells, human umbilical cord perivascular cells (HUCPVCs), adipose stem cells, and liver stem cells.
  • IPS induced pluripotent stem cells
  • IPS hematopoietic stem cells
  • intestinal stem cells intestinal stem cells
  • osteoblastic stem cells mesenchymal stem cells
  • MPCs multipotent adult progenitor cells
  • mesenchymal stem cells are selected from lung mesenchymal stem cells, Wharton's Jelly mesenchymal stem cells, bone marrow mesenchymal cells, bone marrow stromal cells, umbilical cord mesenchymal cells, spleen mesenchymal cells (e.g., Hox1 1 + , CD45 " cells), adipose derived mesenchymal cells, and pericytes.
  • the epithelial stem cells are selected from the group consisting of lung epithelial stem cells, breast epithelial stem cells, vascular epithelial stem cells, and intestinal epithelial stem cells.
  • the skin stem cells are selected from the group consisting of epidermal stem cells and follicular stem cells (hair follicle stem cells).
  • the neural cells are selected from neuronal dopaminergic stem cells and motor neuron stem cells.
  • the cell therapy apparatus or device could be used with any or all of these so named cells (or other cells) to perform similar scientific, therapeutic, or industrial uses.
  • Each of these cell types could be immortalized to provide for an unlimited supply of progenitor cells for use in the cell therapy apparatus or device.
  • the cell therapy apparatus or device containing immortalized progenitor or stem cells, such as MSCs, or those cells and cell types described above that provide repair and regeneration factors may also be useful to study recovery from cerebral stroke, traumatic brain injury, neural injury, or spinal cord injury (SCI), and to provide new therapeutic or industrial uses (Yasuhara et al., J. Cereb. Blood Flow Metab. 28:1804-1810, 2008; Chopp et al., Lancet Neurol.
  • Immortalized cells are cells that have the capacity to divide and proliferate indefinitely either in vivo or in vitro over a long period of time.
  • Techniques for immortalizing cells are described in WO 2003014320, US 5716830, US 5648219, WO 2013147082, WO2013186264, WO 2002059285, US 20120237607, WO 2000061617, WO 1997032992, WO 1997023602, US 7776587, and WO20131 18786, which are incorporated herein by reference in their entireties.
  • methods of making immortalized cells generally include introducing an "immortalizing gene,” such as an oncogene, a cell proliferation factor gene, a telomerase gene to the cells using virus-based cell transduction methods.
  • an "immortalizing gene” such as an oncogene, a cell proliferation factor gene, a telomerase gene
  • Other transduction methods are also available in the art, such as electroporation, liposome-based gene delivery, or Ca 2+ phosphate precipitation (see, e.g., Wigler et al., Cell 16:777, 1979), but virus-based transduction methods generally give higher transduction efficiency. Normally, the following transduction methods are also available in the art, such as electroporation, liposome-based gene delivery, or Ca 2+ phosphate precipitation (see, e.g., Wigler et al., Cell 16:777, 1979), but virus-based transduction methods generally give higher transduction efficiency. Normally,
  • immortalizing genes and viruses used for transduction are first packaged and secreted into cell culture medium by other cells that are used particularly for the purpose of generating these viruses.
  • the packaged viruses containing an "immortalizing gene" in the culture medium are collected and added to cells to inoculate them.
  • viral mediated transduction methods are available in the art, such as adenovirus- mediated transduction, retrovirus-mediated transduction, adeno-associated virus-mediated transduction, lentivirus-mediated transduction, and herpes virus-mediated transduction.
  • the immortalizing genes introduced may or may not be under the control of an inducible promoter.
  • the genes Once the genes are incorporated into the genome of the cell, they can be transmitted to the daughter cells and therefore can be expressed stably over a long period of time.
  • the oncogene or telomerase gene would be able to provide cells with the ability to proliferate indefinitely. After the cells are successfully transduced with immortalizing genes, they are often cultured in the presence of a combination of growth factors to ensure that the introduced gene is active to enable the production of immortalized cells.
  • immortalized stem cells e.g., im-hMSCs
  • T antigen a proto-oncogene
  • the T antigen is constitutively expressed, and the cells grow robustly.
  • an inducible Tetracycline-on large T antigen vector can be used such that the stem cells (e.g., hMSCs) are cultured and produced with doxycycline (dox) present in the culture medium.
  • dox doxycycline
  • Tetracycline-on promoters and vectors expressing T antigen are well known in the field.
  • Another method for immortalizing stem cells e.g., hMSCs, utilizes the ecdysone receptor-based inducible gene expression system for promoting expression of the immortalizing gene (see, e.g., U.S. Patent No. 7,776,587, which is incorporated by reference herein).
  • stem cells in addition to differentiating into cells of specialized tissues or organs, stem cells also produce and secrete useful therapeutic substances in the form of cytokines, chemokines, growth factors, or anti-inflammatory agents that can influence their neighboring cells.
  • useful therapeutic substances in the form of cytokines, chemokines, growth factors, or anti-inflammatory agents that can influence their neighboring cells.
  • some stem cells are not static producers of these substances but respond to the molecules and the chemical environment nearby to modulate their production of certain useful substances.
  • the apparatus or device of the invention transfers the therapeutic substances or bioactive factors produced by the resident cells in the apparatus or device to the recipient damaged organs or tissues.
  • Bioactive factors produced by mesenchymal stem cells include interleukins (IL-) 1 a, 1 b, 6, 7, 8, 10, 1 1 , 14 and 15, macrophage colony stimulating factor (M-CSF), granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), stem cell factor (SCF-1 ), leukemia inhibitory factor (LIF), prostaglandin E-2 (PGE-2), transforming growth factor ⁇ (TGF- ⁇ ), hepatocyte growth factor (HGF), stromal cell derived factor-1 (SDF-1 ), thrombopoetin (TPO), Flt-3 ligand, exosomes containing factors, pro-factors, mRNAs, and immune modulating factors including TGF- ⁇ , IL-1 a receptor antagonist (IL-1 Rag), indoleamine 2,3-dioxygenase (IDO), inducible nitric oxide synthase (iNOS), galectin 1
  • the hMSCs are known to produce up to 20 factors that are useful in repairing tissues and organs.
  • factors produced by hMSCs that inhibit inflammation include TGF- ⁇ , HGF, PGE-2, Gal-1 , iNOS, IL-6, CD73, IL-1 Rag, IL-10, HLA-G, IDO, and TSG-6.
  • Growth factors produced by hMSCs include M-CSF, G-CSF, GM-CSF, LIF, SCF, Flt-3
  • exosomes from hMSCs which contain factors travel long distances in the body. At least 1 1 factors produced by hMSCs are known to reduce inflammatory responses (see FIG. 5, and Aggarwal and Pittenger Blood 105:1815-1822, 2005; Pittenger et al., Cell Stem Cell. 5:8-10, 2009).
  • hMSCs also interact with proinflammatory dendritic cells (DC1 ) to reduce production of TNFa and IL-12, and with anti-inflammatory dendritic cells (DC2) to increase IL-10 and IL-4.
  • DC1 proinflammatory dendritic cells
  • DC2 anti-inflammatory dendritic cells
  • hMSCs interact with proinflammatory T cells (T H 1 ) to reduce expression of interferon Y, and with anti-inflammatory T cells (T H 2) to increase expression of IL-4 and IL-5.
  • T H 1 proinflammatory T cells
  • T H 2 anti-inflammatory T cells
  • antiinflammatory Tregs are also increased by hMSCs.
  • similar effects are seen in vivo on T cell activity and inflammation, and injection of hMSCs at 1 -2 million per kg body weight are effective in treating severe GvHD grade III and normally deadly grade IV GvHD (Le Blanc et al., Lancet 363:1439-41 , 2004).
  • Tissues and organs that can benefit from factors produced by immortalized stem or progenitor cells and provided by the cell therapy apparatus or device of the invention are provided by the cell therapy apparatus or device of the invention.
  • immortalized stem or progenitor cells such as hMSCs, and other useful cells can produce many bioactive factors that are able to provide therapeutic benefits.
  • the apparatus or device of the invention provides a way to supply those factors to tissues or organs in need.
  • the organs that may benefit from the apparatus or device include, but are not limited to, lung, heart, liver, bladder, brain, blood vessels, skin, eye structures, gut, bone, muscle, ligament, cartilage, esophagus, pancreas, intestines, gallbladder, bile duct, fallopian tubes, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureter, urethra, uterus, or fat.
  • the tissues that may benefit from the apparatus or device include, but are not limited to, tissues of the lung, heart, liver, bladder, brain, blood vessels, nervous system, skin, eye structures, gut, bone, muscle, ligament, cartilage, esophagus, pancreas, intestines, gallbladder, bile duct, fallopian tubes, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureter, urethra, uterus, and fat.
  • the apparatus or device of the invention is especially useful in providing benefits to organs that are undergoing transplant surgeries.
  • the apparatus or device may be attached to the donor or recipient extracorporeally, proximal to the organ of interest in the case of the donor.
  • the apparatus or device may also be attached to an organ that is already harvested from a donor and is being transported to a recipient.
  • the apparatus or device may also be connected to the recipient after transplant surgeries. In all cases, the apparatus or device would be able to transfer therapeutic bioactive factors to the organ or recipient and could be used to improve transplant outcomes.
  • MSCs or other stem cells used in the apparatus or device containing immortalized cells can be powerful producers of growth factors and cytokines, and of factors that modulate immune responses including PGE-2, TGF- ⁇ , IL-1 a receptor antagonist (IL-1 Rag), indoleamine 2,3-dioxygenase (IDO), inducible nitric oxide synthase (iNOS), galectin 1 (Gal-1 ), human leukocyte antigen G (HLA-G), TNF-a stimulated gene/protein 6 (TSG-6), and antibacterial factors PGE-2 and peptide LL-37.
  • graft versus host disease graft versus host disease
  • the newly grafted stem cells produce immune cells that see the host as foreign tissue and attack.
  • the graft produces dendritic cells, T cells, and B cells that conspire to destroy the "foreign" cells around them, and results in tissue damage that can be life threatening if it becomes severe (grade III or grade IV) GVHD.
  • the inflammation, edema, and fever from tissue damage result in additional responses from the body including the profound increase in the release of cytokines that stimulate many responses. This cytokine storm is difficult for clinicians and nurses to manage as so many biochemical and tissue reactions occur simultaneously.
  • the apparatus or device containing immortalized cells can be used to modulate the immune response that occurs in GVHD and subsequent cytokine storm(s).
  • cancer researchers have learned to harness the immune response to attack cancer cells in the body. This is done by isolating a patient's dendritic cells and exposing them to known cancer antigens, which they engulf, process, and present to T cells and B cells to stimulate the immune response against the cancer antigen. When this works properly it can result in the rapid elimination of cancer cells that express the known cancer antigen from the body. Such patient treatment can be highly successful. However, the accelerated immune response also can result in life threatening cytokine storm that causes rapid and painful physiological changes that are life threatening. All the biochemical and tissue reactions occur rapidly and are difficult for the clinicians/nurses to effectively treat with current therapies. A patient's condition may quickly go from stable to life threatening within tens of minutes.
  • the apparatus or device containing immortalized cells such as immortalized MSCs or other stem/progenitor cells which releases factors that modulate the immune response can be effective in preventing or alleviating life threatening cytokine storm.
  • immortalized cells such as immortalized MSCs or other stem/progenitor cells which releases factors that modulate the immune response
  • the physician has another tool to intervene in a situation where there are few choices.
  • the apparatus or device of the invention can also provide a method of treatment for various medical conditions which include, but are not limited to, an acute disease, a chronic disease, an autoimmune disease, a skin disease, a neurodegenerative disease, a musculoskeletal disease, cancer, a vascular or circulatory disease, an inflammatory condition, a rheumatology disease, cardiac infarction, acute or chronic heart disease, acute or chronic respiratory disease or distress, trauma, sepsis, and organ failure.
  • various medical conditions include, but are not limited to, an acute disease, a chronic disease, an autoimmune disease, a skin disease, a neurodegenerative disease, a musculoskeletal disease, cancer, a vascular or circulatory disease, an inflammatory condition, a rheumatology disease, cardiac infarction, acute or chronic heart disease, acute or chronic respiratory disease or distress, trauma, sepsis, and organ failure.
  • Cardiomyopathy Celiac Sprue-Dermatitis, Chronic Fatigue Immune Dysfunction Syndrome (CFIDS), Chronic Inflammatory Demyelinating Polyneuropathy, Churg-Strauss Syndrome, Cicatricial Pemphigoid, Limited Scleroderma (CREST Syndrome), Cold Agglutinin Disease, Crohn's Disease, Discoid Lupus, Essential Mixed Cryoglobulinemia, Fibromyalgia-Fibromyositis, Graves' Disease, Guillain-Barre
  • IgA Thrombocytopenia Purpura
  • IgA IgA Nephropathy, Insulin dependent Diabetes, Juvenile Arthritis, Lichen Planus, Lupus, Meniere's Disease, Mixed Connective Tissue Disease, Multiple Sclerosis, Myasthenia Gravis, Pemphigus Vulgaris, Pernicious Anemia, Polyarteritis Nodosa, Polychondritis, Polyglandular Syndromes, Polymyalgia Rheumatica, Polymyositis and Dermatomyositis, Primary Agammaglobulinemia, Primary Biliary Cirrhosis, Psoriasis, Raynaud's Phenomenon, Reiter's Syn irome, Rheumatic Fever, Rheumatoid Arthritis, Sarcoidosis, Scleroderma, Sjogren's Syndrome, Stiff-Man Syndrome, Takayasu Arteritis, Temporal Arteritis/Giant Cell Arteritis,
  • the autoimmune disease to be treated is type I diabetes.
  • hMSCs are known to make at least 1 1 factors that modulate inflammation including TGF- ⁇ , PGE-2, HGF, Gal-1 , IL-6, IL-10, IL-1 Rag, HLA-G, IDO and TSG-6. Each factor can reduce inflammation and the combination of factors can be even more potent, making hMSCs highly hopeful in treating severe or refractory inflammatory and/or autoimmune disorders.
  • the use of the cell therapy apparatus or device containing cells that produce these factors will provide a safer method of treatment since it does not require injection of the cells into the patient. Moreover, the cells will remain viable in the apparatus or device for a longer period of time than when delivered into the body.
  • neurodegenerative diseases that may benefit from the invention include, but are not limited to, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS),
  • Huntington's disease cerebral stroke, traumatic brain injury, and spinal cord injury.
  • stem cells from the bone marrow when transplanted into the brain may be effective in slowing the progression of Parkinson's disease.
  • Mesenchymal stem cells produce TGF- ⁇ and monocyte chemoattractant protein 1 (MCP-1 ) and other factors supporting central nervous system cells.
  • MCP-1 monocyte chemoattractant protein 1
  • MCP-1 monocyte chemoattractant protein 1
  • BDNF brain derived neurotrophic factor
  • mesenchymal stem cells are harvested from the bone marrow, cultivated so they multiply, and then infused into the arteries that supply blood to the brains of patients.
  • Arterial delivery of cells which are larger than most capillaries, can result in ischemia and stroke risks.
  • bioactive factors as provided by the cell therapy apparatus or device containing MSCs or other
  • stem/progenitor cells and not cell infusion can limit such risks.
  • mesenchymal stem cells are being tested in patients with ALS at several doses given intrathecally (spinal cord delivery). Recipients will receive injection of 10 million cells, 50 million cells, 100 million cells or 2 doses of 50 million or 100 million cells 4 weeks apart. Owing to their production of anti-inflammatory molecules and growth factors, it is envisioned that the cells will improve the clinical outcome.
  • the protocol requires 8 weeks for the isolation and proliferation of the MSCs from the patient before treatment can begin.
  • immortalized cells can begin immediately as no waiting period for cell growth is needed.
  • the PROCHYMAL ® cell product (Osiris Therapeutics, Inc.) could be incorporated in the apparatus or device of the invention.
  • This apparatus or device could be used in the treatment of GvHD, Crohn's Disease, chronic obstructive pulmonary disease (COPD), acute myocardial infarction, diabetes, and acute radiation syndrome (ARS) in a subject (or organ or tissue) in need.
  • COPD chronic obstructive pulmonary disease
  • ARS acute radiation syndrome
  • CHONDROGEN ® Osiris Therapeutics, Inc.
  • This apparatus or device could be used in the treatment of osteoarthritis.
  • the MULTISTEM ® cell product (Athersys, Inc.) could be incorporated in the apparatus or device of the invention.
  • This apparatus or device could be used in the treatment of 1 ) inflammatory and immune diseases, such as diabetes, allergies, rheumatoid arthritis (RA), immune deficiency disorders, inflammatory bowel disease (e.g., IBD, Crohn's, ulcerative colitis), lupus, graft- versus-host disease (GvHD), dermatological conditions (e.g., psoriasis, eczema), pelvic inflammatory disease, pulmonary conditions, scleroderma, transplant rejection, vasculitis, and a range of others; 2) neurological diseases and disorders, such as acute trauma or ischemic injury conditions, such as stroke (e.g., ischemic stroke), traumatic brain injury (TBI), spinal cord injury, and neonatal hypoxic ischemia, as well as episodic, chronic, or progressive neurological conditions, such as depression, schizophrenia, epit
  • PDA-002 (Celgene, Inc.) could be incorporated into an apparatus or device of the invention.
  • This apparatus or device could be used in the treatment of, e.g., peripheral artery disease and diabetic foot ulcers in diabetic patients, or for other recognized uses to which the PDA-002 cell therapy product has been applied.
  • PLX-PAD Pluristem, Inc
  • This apparatus or device could be used in the treatment of, e.g., intermittent claudication (Charcot's Syndrome), which is related to peripheral artery disease, or for other recognized uses to which the PLX-PAD cell therapy product has been applied.
  • intermittent claudication Charge's Syndrome
  • mesenchymal progenitor cells (MPC) (Mesoblast Ltd.) could be incorporated into an apparatus or device of the invention.
  • This apparatus or device could be used in the treatment of, e.g., myocardial infarction, or for other recognized uses to which the MPC cell therapy product has been applied.
  • ALD-401 , ALD-301 , ALD-201 , ALD-151 , and ALD-601 cells could be incorporated into an apparatus or device of the invention.
  • An apparatus or device of the invention containing ALD-401 cells could be used in the treatment of, e.g., stroke, or for other recognized uses to which the ALD-401 cell therapy product has been applied.
  • An apparatus or device of the invention containing ALD-301 cells could be used in the treatment of, e.g., critical limb ischemia, or for other recognized uses to which the ALD-301 cell therapy product has been applied.
  • An apparatus or device of the invention containing ALD-201 cells could be used in the treatment of, e.g., ischemic heart failure, or for other recognized uses to which the ALD-201 cell therapy product has been applied.
  • An apparatus or device of the invention containing ALD-151 cells could be used, e.g., to improve engraftment following cord blood transplants in the treatment of leukemia, or for other recognized uses to which the ALD-151 cell therapy product has been applied.
  • An apparatus or device of the invention containing ALD- 601 cells could be used in the treatment of, e.g., inherited metabolic diseases, or for other recognized uses to which the ALD-601 cell therapy product has been applied.
  • any of the diseases and disorders discussed above in connection with any specific cell type could be treated using an apparatus or device of the invention that contains any of the other disclosed cell types in the application.
  • an apparatus or device of the invention that contains one or more of the cell types disclosed herein could be used in the treatment of one or more of the following: 1 ) inflammatory and immune diseases, such as diabetes, allergies, rheumatoid arthritis (RA), immune deficiency disorders, inflammatory bowel disease (e.g., IBD, Crohn's, ulcerative colitis), lupus, graft-versus-host disease (GvHD), dermatological conditions (e.g., psoriasis, eczema), pelvic inflammatory disease, pulmonary conditions, scleroderma, transplant rejection, vasculitis, cytokine storm, and a range of others; 2) neurological diseases and disorders, such as acute trauma or ischemic injury conditions, such as stroke (e.g., ischemic stroke), traumatic brain injury (TB).
  • MSCs e.g., bone marrow MSCs
  • CABG coronary artery bypass grafting
  • the patients receive 10-20 needle injections into the healthy areas of the heart to deliver 20 million to 200 million MSCs.
  • the use of the cell therapy apparatus or device of the invention would eliminate the waiting period for the growth of the cells, and avoid the many needle sticks into the healthy heart tissue and subsequent risks.
  • acute respiratory distress syndrome can be caused by many different types of lung injury and the injury causes inflammation.
  • the many anti-inflammatory factors produced by immortalized hMSCs or other stem/progenitor cells can provide reparative effects in the lungs and limit fibrosis and other impairment.
  • Treatment doses for infused MSCs are currently 1 , 5 and 10 million per kilogram of body weight (up to 700 million for a typical 70 kg man).
  • the cell therapy apparatus or device of the invention would provide the same potential benefit and without the risks of injecting the cells into the patient.
  • the cell therapy apparatus or device of the invention may be used to improve the transplant outcomes of organ transplants, including lung, heart, kidney, and liver transplants.
  • the apparatus or device may be connected to the in vivo perfused organ prior to organ harvest from the donor.
  • the apparatus or device may be connected to the ex vivo perfused organ after the organ is harvested from the donor, but before it is transplanted to the recipient.
  • the apparatus or device is connected to the ex vivo perfused organ during organ transport between donor and recipient to improve transplant outcome.
  • the cell therapy apparatus or device of the invention may be used to improve the outcome following trauma-related acute respiratory distress syndrome, such as that which accompanies chest damage related to motor vehicle accidents.
  • the tissue injury may be sterile, but the damage causes an inflammatory response by the body.
  • the device or apparatus of the invention containing immortalized hMSCs can be connected to the circulatory system to provide anti-inflammatory factors and improve outcome.
  • the cell therapy apparatus or device containing immortalized hMSCs may be used to reduce bacterial sepsis by delivery of antibacterial prostaglandin E2 (PGE2) and LL-37 peptides.
  • PGE2 antibacterial prostaglandin E2
  • LL-37 peptides
  • MSCs do not increase bacterial sepsis.
  • a device or apparatus of the invention containing immortalized hMSCs can be connected to the circulatory system of a patient having or suspected of having bacterial sepsis to provide antibacterial factors and improve outcome.
  • the apparatus or device containing the immortalized cells can be used to produce conditioned medium that has therapeutic value.
  • the medium is collected after passage through the apparatus and this conditioned medium can be used as is, concentrated, or combined with other factors known to be beneficial.
  • the immortalized cells in the apparatus may be treated with one or more chemicals, bioactive factors, or other cell types that alter their production of molecules from the resident, immortalizd cells such that the conditioned medium now has a new composition.
  • another cell type such as peripheral blood mononuclear cells are added to the apparatus.
  • the peripheral blood mononuclear cells cause the resident, immortalized cells of the apparatus to produce much more VEGF (vascular endothelial growth factor).
  • VEGF vascular endothelial growth factor
  • the bioactive factor such as TGFfi
  • TGFfi is inoculated into the apparatus containing the immortalized cells and allowed to interact with the cells for 1 to 36 hrs and the conditioned medium is collected and used for therapeutic purposes or analyzed (see Example 8) to identify the novel molecules produced by the treated cells.
  • the apparatus or device containing the immortalized cells can also be implanted to provide the same benefits or a treatment that may extend over a longer period of time - days, weeks, months, or years. Implantation also provides for an optimal temperature for maintaining the cells in the apparatus or device. For example, the recovery from a traumatic injury may occur over weeks and the implantation of a device or apparatus containing immortalized cells may provide extended benefits not possible with a brief treatment. Further, the cells in the device can be bioengineered to express additional molecules that may provide benefits to specific patients such as diabetic patients, wherein the cells of the apparatus would be engineered to express insulin of a beneficial amount.
  • the apparatus or device containing the cells allows the cell-produced molecules to be sampled over time or when the cells in the apparatus are treated with another bioactive factor such as TNFa (Perdoni et al. Stem Cell Research & Therapy 5:121 , 2014; Beltran et al, J Pehodontol. 86:62-71 , 2015) or TGFfi, or other cells are inoculated into the apparatus such as endothelial cells to study molecules produced following cell-cell interactions.
  • TNFa Perdoni et al. Stem Cell Research & Therapy 5:121 , 2014; Beltran et al, J Pehodontol. 86:62-71 , 2015
  • TGFfi TGFfi
  • the apparatus or device containing the immortalized cells can be used to determine which produced molecules are therapeutically effective. For this, the conditioned medium from the cells in the device are analyzed by separation with gas chromatography followed by mass spectrometry for identifying the individual molecules. The individual molecules can then be removed from the original conditioned medium by chemical means or by immuno-depletion. After depletion of individual molecules, the remaining molecules can be tested in vitro or in vivo for therapeutic value. By using this approach iteratively, the important active molecules can be determined. Knowing these active molecules, the parameters of the apparatus containing the immortalized cells can be optimized to provide more of these molecules by methods known to one knowledgeable in the field.

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Abstract

L'invention concerne un appareil ou un dispositif pour une thérapie cellulaire extracorporelle, implantée ou ex vivo, des procédés d'utilisation de l'appareil ou dispositif pour traiter divers états médicaux, tels qu'une maladie auto-immune, une maladie neurodégénérative, un cancer, des états inflammatoires, une crise cardiaque, une maladie cardiaque chronique, une détresse respiratoire aiguë, un traumatisme, une défaillance organique, et un kit contenant l'appareil ou le dispositif. L'appareil ou dispositif peut être relié à un sujet, organe ou tissu et peut être utilisé pour transférer des substances thérapeutiques, produites par des cellules immortelles qui résident dans l'appareil ou dispositif, au sujet, organe ou tissu.
PCT/US2015/025973 2014-04-15 2015-04-15 Appareil ou dispositif pour thérapie cellulaire WO2015160952A1 (fr)

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CN110241073A (zh) * 2019-07-15 2019-09-17 朱家源 快速分离提取表皮干细胞的方法
WO2020097449A1 (fr) * 2018-11-09 2020-05-14 Figene, Llc Moyens et méthodes de prévention ou d'inversion du vieillissement
CN115725686A (zh) * 2021-08-30 2023-03-03 四川大学 基于小电导机械力敏感性通道的生物埃米孔系统

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WO2020097449A1 (fr) * 2018-11-09 2020-05-14 Figene, Llc Moyens et méthodes de prévention ou d'inversion du vieillissement
JP2022512953A (ja) * 2018-11-09 2022-02-07 フィジーン、エルエルシー 老化を防止又は逆転させる手段及び方法
CN110241073A (zh) * 2019-07-15 2019-09-17 朱家源 快速分离提取表皮干细胞的方法
CN110241073B (zh) * 2019-07-15 2021-07-13 广州市麦施缔医疗科技有限公司 快速分离提取表皮干细胞的方法
CN115725686A (zh) * 2021-08-30 2023-03-03 四川大学 基于小电导机械力敏感性通道的生物埃米孔系统
CN115725708A (zh) * 2021-08-30 2023-03-03 四川大学 一种利用生物埃米孔检测核苷酸的方法
CN115927738A (zh) * 2021-08-30 2023-04-07 四川大学 一种检测样本中目标核酸的存在的方法

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