Mixed cell populations for tissue repair and separation technique for cell processing

• US 9,415,071 B2
• Filed: 03/04/2013
• Issued: 08/16/2016
• Est. Priority Date: 11/03/2006
• Status: Active Grant

• Current Assignee: Vericel Corporation
• Original Assignee: Vericel Corporation
• Inventors: Hampson, Brian, Goltry, Kristin, Smith, Douglas M., Rowley, Jonathan A., Venturi, Naia
• Primary Examiner(s): FOX, ALLISON M
• Application Number:
  US13/784,066
• Publication Number:
  US 20130209420A1
• Time in Patent Office: 1,261 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61K 39/461 characterised by the cell type used
• CPC Class Codes: A61K 39/464 characterised by the antigen targeted or presented
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/30 : Organic components
• CPC Class Codes: C12N 2500/32 : Amino acids
• CPC Class Codes: C12N 2500/84 : from mammals
• CPC Class Codes: C12N 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 5/0645 : Macrophages, e.g. Kuepfer cells in the liver; Monocytes
• CPC Class Codes: C12N 5/0663 : Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0669 : Bone marrow stromal cells; Whole bone marrow isolated stem cells from bone marrow C12N5/0647, C12N5/0663

Abstract

The present invention provides a fluid exchange cell culture technique and tissue repair cells (TRCs) made by these methods, as well as methods using these cells. The method includes a new wash step which increases the tissue repair properties of the TRCs of the invention. This wash step allows for the production of TRC populations with greater tissue repair and anti-inflammatory capabilities. Embodiments of the present invention include a post-culture process for cultured cells that preferably includes the steps of: a wash process for removing unwanted residual culture components, a volume reduction process, and a harvesting process to remove cultured cells. Preferably, all these steps are performed within a aseptically closed cell culture chamber by implementing a separation method that minimizes mechanical disruption of the cells and is simple to automate. The harvested cells may then be concentrated to a final volume for the intended use. In such embodiments, the final composition is a substantially purified and concentrated cell mixture suspended in a physiologic solution suitable for immediate use in humans without further washing, volume reduction, or processing. Embodiments are also applicable to harvesting (and/or washing) particles within a liquid or solution within a chamber.

Claims

1. A composition for tissue repair comprising

a bio-compatible matrix comprising a synthetic biodegradable polymeric material selected from the group consisting of poly-lactides, poly-glycolides, poly-fumarates and poly-ethylene glycol,

and

a mixed population of cells of hematopoietic, mesenchymal and endothelial lineage,

wherein the viability of said cells is at least 80% and the composition contains:

a) about 5-75% viable CD90⁺ cells with the remaining cells in said composition being CD45⁺;

b) less than 2 μ

g/ml of bovine serum albumin;

c) less than 1 μ

g/ml of a enzymatically active harvest reagent;

wherein the cells produce

at least one anti-inflammatory cytokine selected from the group consisting of interleukin-1 receptor antagonist, interleukin-6, TGF-β

, interleukin-8, interleukin 10, and monocyte chemoattractant protein-1, or

at least one angiogenic factor; and

wherein the composition is further characterized as being substantially free of mycoplasm, endotoxin, and microbial contamination.

2. The composition of claim 1, wherein the cells are derived from mononuclear cells.

3. The composition of claim 2, wherein the mononuclear cells are derived from bone marrow, peripheral blood, umbilical cord blood or fetal liver.

4. The composition of claim 1, wherein said cells are in a pharmaceutical-grade electrolyte solution suitable for human administration.

5. The cell composition of claim 1, wherein said angiogenic factor is vascular endothelial growth factor, angiopoeitin 1, angiopoeitin 2 or hepatocyte growth factor.

6. The cell composition of claim 1, wherein said cells produce less than 10 pg/mL per 24 hour period per 10⁵ cells of one or more pro-inflammatory cytokines.

7. The cell composition of claim 6, wherein said pro-inflammatory cytokine is interleukin-1 alpha, interleukin-1 beta, interferon gamma or interleukin-12.

8. The cell composition of claim 1, wherein the cells expresses indoleamine 2,3, dioxygenase or PD-L1.

9. The composition of claim 1, wherein at least 10% of said CD90⁺ co-express CD15.

10. The composition of claim 1, wherein said CD45⁺cells are CD14⁺, CD34⁺ or VEGFR1⁺.

11. The composition of claim 1, wherein said composition is substantially free of horse serum and/or fetal bovine serum.

12. The composition of claim 1, wherein the total number of viable cells is 35 million to 300 million.

13. The composition of claim 12, wherein said cells are in a volume less than 15 milliliters.

14. The composition of claim 12, wherein said cells are in a volume less than 10 milliliters.

15. The composition of claim 12, wherein said cells are in a volume less than 7.5 milliliters.

16. The composition of claim 1, wherein the composition is useful in treating pathologies selected from the group consisting of ischemic conditions, conditions requiring organ or tissue regeneration, inflammatory diseases and auto-immune diseases.

17. The composition of claim 16, wherein the ischemic conditions are selected from the group consisting of limb ischemia, congestive heart failure, cardiac ischemia, kidney ischemia, ESRD, stroke, and ischemia of the eye.

18. The composition of claim 16, wherein the organ or tissue regeneration is selected from the group consisting of regeneration of liver, pancreas, lung, salivary gland, blood vessel, bone, skin, cartilage, tendon, ligament, brain, hair, kidney, muscle, cardiac muscle, nerve, and limb.

19. The composition of claim 16, wherein the inflammatory disease is selected from the group consisting of heart disease, diabetes, spinal cord injury, rheumatoid arthritis, osteo-arthritis, inflammation due to hip replacement or revision, Crohn'"'"'s disease, and graft versus host disease.

20. The composition of claim 16, wherein the auto-immune disease is selected from the group consisting of type 1 diabetes, psoriasis, systemic lupus, and multiple sclerosis.

21. A tissue repair reagent comprising the composition of claim 1.