Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/14—Blood; Artificial blood
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/06—Animal cells or tissues; Human cells or tissues
  • C12N5/0602—Vertebrate cells
  • C12N5/0634—Cells from the blood or the immune system
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/06—Animal cells or tissues; Human cells or tissues
  • C12N5/0602—Vertebrate cells
  • C12N5/0634—Cells from the blood or the immune system
  • C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
  • C12N2501/10—Growth factors
  • C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
  • C12N2501/20—Cytokines; Chemokines
  • C12N2501/22—Colony 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)

Priority Applications (1)

Applications Claiming Priority (1)

Related Child Applications (1)

Publications (1)

Family

ID=34421167

Family Applications (2)

Family Applications Before (1)

Country Status (7)

Families Citing this family (1)

Family Cites Families (5)

• 2003
  • 2003-09-02 EP EP08007867A patent/EP1947173A3/de not_active Withdrawn
  • 2003-09-02 CN CNA038270188A patent/CN1826405A/zh active Pending
  • 2003-09-02 BR BRPI0318490-0A patent/BR0318490A/pt not_active IP Right Cessation
  • 2003-09-02 AU AU2003265878A patent/AU2003265878A1/en not_active Abandoned
  • 2003-09-02 EP EP03818826A patent/EP1660643A1/de not_active Withdrawn
  • 2003-09-02 JP JP2005509358A patent/JP2007520993A/ja active Pending
  • 2003-09-02 WO PCT/US2003/027398 patent/WO2005033299A1/en active Application Filing
  • 2003-09-02 CA CA002537318A patent/CA2537318A1/en not_active Abandoned

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events