EP1660643A1 - Procede de reparation d'un tissu de mammifere primate - Google Patents

Procede de reparation d'un tissu de mammifere primate

Info

Publication number
EP1660643A1
EP1660643A1 EP03818826A EP03818826A EP1660643A1 EP 1660643 A1 EP1660643 A1 EP 1660643A1 EP 03818826 A EP03818826 A EP 03818826A EP 03818826 A EP03818826 A EP 03818826A EP 1660643 A1 EP1660643 A1 EP 1660643A1
Authority
EP
European Patent Office
Prior art keywords
tissue
cell
repaired
cells
blood cells
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP03818826A
Other languages
German (de)
English (en)
Inventor
Donnie Rudd
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Regenetech Inc
Original Assignee
Regenetech 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 Regenetech Inc filed Critical Regenetech Inc
Priority to EP08007867A priority Critical patent/EP1947173A3/fr
Publication of EP1660643A1 publication Critical patent/EP1660643A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14Blood; Artificial blood
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0634Cells from the blood or the immune system
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0634Cells from the blood or the immune system
    • C12N5/0647Haematopoietic stem cells; Uncommitted or multipotent progenitors
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10Growth factors
    • C12N2501/125Stem cell factor [SCF], c-kit ligand [KL]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20Cytokines; Chemokines
    • C12N2501/22Colony stimulating factors (G-CSF, GM-CSF)

Definitions

  • the present invention relates to a method of repairing primate mammalian tissue.
  • Regeneration of primate mammalian tissue has long been a desire of the medical community.
  • repair of primate mammalian tissue has been accomplished largely by transplantations of like tissue from a donor.
  • tissue transplantation began essentially with the kidney transplant from one of the Herrick twins to the other and later made world famous by South Af ican Doctor Christian Barnard's transplant of a heart from Denise Darval to Louis Washkansky on December 3, 1967, tissue transplantation became a widely accepted method of extending life in terminal patients.
  • Transplantation of mammalian tissue, from its first use encountered major problems, primarily tissue rejection due to the body's natural immune system.
  • tissue stem cells such as the transplantation of liver stem cells found in U.S. Patent No. 6,129,911 have similar limitations rendering their widespread use questionable.
  • researchers have experimented with the use of pluripotent embryonic stem cells as an alternative to tissue transplant
  • the theory behind the use of embryonic stem cells has been that they can theoretically be utilized to regenerate virtually any tissue in the body.
  • the use of embryonic stem cells for tissue regeneration has also encountered problems.
  • the present invention is a method of repairing primate mammalian tissue.
  • the method comprises removing blood cells from the primate mammal, controllably expanding the blood cells by a factor of at least seven times preferably in less than seven days while ma taining their three-dimensional geometry and their cell-to- cell support and cell-to-cell geometry, and reintroducing the expanded blood cells into the primate mammal within a time period sufficient to allow the primate mammal body system to utilize the blood cells to effectively repair damaged tissue. It is an object of this invention to provide a method of r airing primate mammalian tissue. It is a further object of this invention to use a combination of expanded blood and the body's ability to repair itself to repair body tissue.
  • the present invention is related to a method of repairing, replenishing and regenerating tissue in primate mammalians.
  • This invention may be more fully described by the preferred embodiment as hereinafter described, but is not intended to be limited thereto.
  • a method is described to prepare adult stem cells that can assist the body in repairing, replacing and regenerating tissue.
  • Blood cells are removed from a patient. A subpopulation of these cells is currently referred to as adult stem cells.
  • the blood cells are placed in a bioreactor such as that described in United States Patent 5,702,941 and is incorporated herein by reference.
  • the bioreactor vessel is rotated at a speed that provides for suspension of the blood cells to maintain their three-dimensional geometry and their cell-to-cell support and geometry.
  • the cells are fed nutrients, exposed to either hormones, cytokines, or growth factors, and/or genetically modified, and toxic materials are removed.
  • the toxic materials typically removed are from blood cells comprising the toxic granular material of dying cells and the toxic material of granulocytes and mycrophages. A subpopulation of these cells is expanded creating a large amount of cells.
  • the expansion of the cells is controlled so that the cells expand at least seven times in a sufficient amount of time, preferably within seven days.
  • the cells are then injected intravenously or directly into the tissue to be repaired allowing the body's natural system to repair and regenerate the tissue.
  • the method can be used to repair liver tissue, heart tissue, hematopoietic tissue, blood vessels, skin tissue, muscle tissue, gut tissue, pancreatic tissue, central nervous system cells, bone, cartilage, connective tissue, pulmonary tissue, spleen tissue, and other body tissue.
  • blood cells can be manipulated to alter their curative characteristics, preferably by genetically modifying the blood cells.
  • peripheral blood (PB) cells are obtained from a person needing tissue repair.
  • MNCs mononuclear cells (MNCs) are obtained from the first apheresis product collected from the donors.
  • the donor Prior to apheresis, the donor is treated with G-CSF 6 :g/kg every 12 hr over 3 days and then once on day 4.
  • MNCs are collected by subjecting the donor's total blood volume to 3 rounds of continuous-flow leukapheresis through a separator, such as a Cobe Spectra cell separator.
  • the present invention relates to a method of replenishing primate mammalian cells. Blood cells are removed by conventional methods from a primate mammal. The cells are contr ⁇ llably expanded in a method utilizing the biosector described in United States Patent 5,702,441, which is incorporated herein by reference. The cells are expanded while mamtaining their three-dimensional geometry and their cell-to- cell geometry.
  • Toxic materials are removed, for example, toxic granular material and mycrophages.
  • the expanded cells are their reintr ⁇ duced by conventional means into the primate animal within a time period that is sufficient to all the primate mammal body system to effectively replenish different cellular population in the primate mammal.
  • Another embodiment of the present invention relates to an ex vivo human blood cell composition that functions to assist a body system to repair, replenish and regenerate tissue, for example, the tissue previously described.
  • the blood cell composition has a substantial number of cells originating from a human hematopoietic system that have undergone ex vivo cell division.
  • the composition comprises at least eight times the. number of blood cells per volume as in the human hermtopoietic system from which it originated.
  • composition is free of toxic granular material, for example, dying cells and the toxic material or content of granulocytes and mychrophages.
  • TMDM Iscove's modified Dulbecco's medium
  • FCS fetal calf serum
  • HA human albumin
  • SCF human stem cell factor
  • Hematopoietic colony-forming cells are assayed using a modification of a previously described assay.
  • 10 5 MNCs are cultured in 0.8% methylcellulose with JJvfDM, 30% FCS, 1.0 U/ml erythropoietin (Amgen), 50 ngml recombinant human GM-CSF (hnmunex Co ., Seattle, WA), and 50 ngml SCF (Amgen). Crce-milliliter aliquots of each culture mixture are then placed in 0.8% methylcellulose with JJvfDM, 30% FCS, 1.0 U/ml erythropoietin (Amgen), 50 ngml recombinant human GM-CSF (hnmunex Co ., Seattle, WA), and 50 ngml SCF (Amgen). Crce-milliliter aliquots of each culture mixture are then placed in
  • Lymphocytes are analyzed by 2-color staining using the following antibody combinations: CD56+CD16-PE/CD3-FTTC, CD3-PE/CD4-FTTC, CD3-PE/CD8- FITC, CD19-PE.
  • Controls include IgGl-PE/IgGl-FLTC for isotype and CD14- PE/CD45-FITC for gating.
  • Progenitor cells are analyzed by 3-color staining with the fluorochromes PerCP/PE FTTC using the following antibody combinations: CD45/CD90/CD34, CD45/CD34/CD38, CD45/CD34/CD33, and CD45/CD34/CD15.
  • CD45/IgGl/IgGl is used as a control.
  • 10 6 cells from the donor are incubated with 10 :1 of antibodies at 2-8EC for 15 minutes in the dark
  • lymphocytes 10,000 cells are acquired from each tube, and then gated on the basis of the forward and right angle light scatter patterns. The cutoff point is visually set at a level above background positivity exhibited by isotype controls.
  • progenitor cells 75,000 cells from each tube is acquired and then sequentially gated.
  • CD34+ cells Incubation of MNCs from normal donors in this tissue culture system significantly increases the numbers of CD34+ cells. The average number of CD34+ cells increased 10-fold by day 6 of culture and plateaus on that same day. The relative number of CD34+ cells co-expressing the myeloid-lineage markers CD15 and CD33 increases significantly by days 5 and 6. After the seventh day, the cells are reinjected into the patient.
  • the injection can be an injection of the cells into the bloodstream or, prererably, an injection directly into the injured tissue such as the liver.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Hematology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Materials For Medical Uses (AREA)

Abstract

L'invention concerne un procédé de réparation d'un tissu de mammifère primate. Ce procédé consiste à prélever des globules sanguins chez un mammifère primate ; à dilater, de manière contrôlée, les globules sanguins par un facteur d'au moins sept fois, de préférence, en moins de sept jours tout en conservant leur géométrie tridimensionnelle, leur support globule-à-globule et leur géométrie cellule-à-cellule, et à réintroduire les globules sanguins dilatés chez le mammifère primate pendant une période suffisante pour permettre au système organisme du mammifère primate d'utiliser les globules sanguins afin de réparer efficacement les tissus endommagés.
EP03818826A 2003-09-02 2003-09-02 Procede de reparation d'un tissu de mammifere primate Withdrawn EP1660643A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP08007867A EP1947173A3 (fr) 2003-09-02 2003-09-02 Procede de preparation des cellules sanguines expansées de mammifere primate

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US2003/027398 WO2005033299A1 (fr) 2003-09-02 2003-09-02 Procede de reparation d'un tissu de mammifere primate

Related Child Applications (1)

Application Number Title Priority Date Filing Date
EP08007867A Division EP1947173A3 (fr) 2003-09-02 2003-09-02 Procede de preparation des cellules sanguines expansées de mammifere primate

Publications (1)

Publication Number Publication Date
EP1660643A1 true EP1660643A1 (fr) 2006-05-31

Family

ID=34421167

Family Applications (2)

Application Number Title Priority Date Filing Date
EP08007867A Withdrawn EP1947173A3 (fr) 2003-09-02 2003-09-02 Procede de preparation des cellules sanguines expansées de mammifere primate
EP03818826A Withdrawn EP1660643A1 (fr) 2003-09-02 2003-09-02 Procede de reparation d'un tissu de mammifere primate

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP08007867A Withdrawn EP1947173A3 (fr) 2003-09-02 2003-09-02 Procede de preparation des cellules sanguines expansées de mammifere primate

Country Status (7)

Country Link
EP (2) EP1947173A3 (fr)
JP (1) JP2007520993A (fr)
CN (1) CN1826405A (fr)
AU (1) AU2003265878A1 (fr)
BR (1) BR0318490A (fr)
CA (1) CA2537318A1 (fr)
WO (1) WO2005033299A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3193893A4 (fr) * 2014-09-16 2018-09-05 Virginia Tech Intellectual Properties, Inc. Compositions de cellules souches, systèmes et utilisations associées

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5702941A (en) 1993-09-09 1997-12-30 Synthecon, Inc. Gas permeable bioreactor and method of use
WO1995021594A1 (fr) 1994-02-09 1995-08-17 Kabi Pharmacia Ophthalmics, Inc. Implantation rapide de lentilles optiques deformables
US5989913A (en) * 1998-07-02 1999-11-23 Charles Daniel Anderson Culture vessel for growing or culturing cells, cellular aggregates, tissues and organoids and methods for using the same
US6129911A (en) 1998-07-10 2000-10-10 Rhode Island Hospital, A Lifespan Partner Liver stem cell
US6541249B2 (en) * 1999-12-22 2003-04-01 Human Genome Sciences, Inc. Immortalized human stromal cell lines

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2005033299A1 *

Also Published As

Publication number Publication date
BR0318490A (pt) 2006-09-12
WO2005033299A1 (fr) 2005-04-14
JP2007520993A (ja) 2007-08-02
AU2003265878A1 (en) 2005-04-21
CN1826405A (zh) 2006-08-30
EP1947173A2 (fr) 2008-07-23
CA2537318A1 (fr) 2005-04-14
EP1947173A3 (fr) 2008-07-30

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