Slide 4 Discover the meaning of “Regenerative Medicine” Stem Cells – What really are they? Learn the sources and types of stem cells Explore the mechanisms of action of stem cells Regenerative Medicine Module Outline
Slide 5 What is Regenerative Medicine? 10 million cells die in your body every minute of every day. Your own stem cells replace them so you can continue living. Goal of Regenerative Medicine: Restitutio In Integrum Restore to Original
Slide 6 Regenerative Medicine Damage Repair Regenerate
Slide 7 Why Use Regenerative Medicine? Current research and clinical trials are exploring regenerative medicine for nearly every organ system. Examples are: Osteoarthritis Tendon/ligament injury Renal failure Liver failure Laminitis Immune-mediated diseases: atopy, IBD, COPD
Slide 8 What are Stem Cells? Pericytes on blood vessels. Courtesy Arnold Caplan and Bruno Peault Stem Cells are: – Primitive cells present in almost every tissue
Slide 9 What are Stem Cells? Stem Cells are: – Primitive cells present in almost every tissue – Able to become different types of tissue: Tendon, Ligament, Bone Stem cells differentiated into cardiomyocytes using growth factors. Courtesy NIH.
Slide 10 What are Stem Cells? Stem Cells are: – Primitive cells present in almost every tissue – Able to become different types of tissue: Tendon, Ligament, Bone Stem cells differentiated into cardiomyocytes using growth factors. Courtesy NIH.
Slide 11 What are Stem Cells? Dividing stem cells. Courtesy Salk Institute. Stem Cells are: – Primitive cells present in almost every tissue – Able to become different types of tissue: Tendon, Ligament, Bone – Self-renewing
Slide 12 What are Stem Cells? Stem Cells are: – Primitive cells present in almost every tissue – Able to become different types of tissue: Tendon, Ligament, Bone – Self-renewing – Pharmaceutical Factories
Slide 13 Definitions Multipotentturn into any cell line of same germ layer Pluripotentturn into any cell line except placental Totipotentturn into any cell type including placental Autograftfrom animal A, into animal A Allograftfrom animal B, into animal A Xenograftfrom species B, into species A Mesenchymal originating from mesoderm
Slide 14 A Rose By Any Other Name Stem cells aka… Mesenchymal stem cells Mesenchymal Stromal cells Multipotent / Pluripotent cells Stromal vascular fraction Nucleated fraction
Slide 15 Embryonic vs Adult Stem Cells Embryonic Source: early embryo ethical dilemma Differentiate into all tissues Purpose: form organism Form Teratomas UNPREDICTABLE Adult Source: all adult tissues (?) no ethical dilemma Differentiate into most tissues Purpose: Regenerate No evidence of Teratoma formation Gruen L and Grabel L, Concise Review: “Scientific and Ethical Roadblocks to Human Embryonic Stem Cell Therapy.” Stem Cell 2006;24;
Slide 16 How to Use Stem Cells Caplan, J Cell. Physiol. 2007, 213: Cell Therapy Injection of non- differentiated cells Cells coordinate healing and regeneration NOW Tissue Engineering Growing tissues and/or organs ex- vivo Stem cells differentiated on a scaffold then implanted Potential Future
Slide 17 Stem Cell Mediated Regeneration 1. Homing (like WBC) 2. Differentiation a. Direct differentiation into needed cell types b. Recruit and stimulate mitosis of local progenitor cells 3. Trophic support - growth factors and cytokines a. Block pain (opioid receptor agonist) b. Down-regulate inflammatory mediators c. Block cell death (anti-apoptosis) d. Stimulate angiogenesis e. Anti-fibrosis (block scar)
Slide 18 Homing Mechanism – Fracture Model Homing of luminescent adipose stem cells to fracture site from IV administration. S-W Lee et al, J Ortho Res, 2009 (Stanford Univ)
Slide 19 Cruciate Ligament - Chondroprotection Toghraie et al, “Treatment of osteoarthritis with infrapatellar fat pad derived mesenchymal stem cells in rabbit” The Knee 2011;1: Sham Surgery ASC IA Control Above toluidine blue staining of cartilage surface at 20 weeks after cranial cruciate ligament transection. Only treatment was group B given 1MM adipose-stem cells by intraarticular injection with no carrier/scaffold.
Slide 20 Eaton M. Cell and molecular approaches to the attenuation of pain after spinal cord injury. J Neurotrauma 2006;23(23/4): Guo – Bone marrow stromal cells produce long-term pain relief in rat models of persistent pain. Stem Cells 2011;29(8): Klass M, Gavrikov V, Csete M et al. Intravenous mononuclear marrow cells reverse neuropathic pain from experimental mononeuropathy. Anesth Analg 2007;104: Malik RA, Veves A, Tesfaye S. Ameliorating human diabetic neuropathy: Lessons from implanting hematopoietic mononuclear cells. Exper Neuro 2006; 201:7-14. Takagi K, Okuda-Ashitaka E, Ito S et al. Involvement of stem cell factor and its receptor tyrosine kinase c-kit in pain regulation. Neurosci 2008;153: Pain Relief Mechanisms
Slide 21 Model – ligation of masseter muscle nerve (constriction injury) Pain Relief Mechanisms Guo et al, Stem Cells 2011;29(8): Normal Highly Sensitized
Slide 22 Anti-inflammation / Anti-fibrosis Co-staining of IL1-RA (red) protein and subpopulation of MSCs (DAPI, blue). Ortiz et al, PNAS (Tulane Univ)
Slide 23 Mechanisms of Regeneration Differentiation into tissue (Photo courtesy Cytori Therapeutics) Nerve Bone Cartilage Liver Fat-derived Stem Cells Cardiac Fat Angiogensis/Anti-apoptosis Gene Therapy Muscle Reviewed in: Tobita M. Adipose-derived stem cells: current findings and future perspectives. Disc Med 2011;11(57):
Slide 24 Mechanisms of Regeneration Stimulation of MSC Proliferation Kol et al (UCD). EVJ 2012.
Slide 25 Mechanisms of Regeneration Stimulation of MSC Migration Kol et al (UCD). EVJ 2012.
Slide 26 Cartilage Regeneration Model Dragoo J et al, “Healing full-thickness cartilage defects using adipose-derived stem cells” Tiss Eng 2007;13(7): (Stanford) At 8 weeks, 12/12 (100%) of defects in treated group healing with hyaline-like cartilage. Only 1/12 (8%) of controls healed.
Slide 27 Anti-apoptosis Mechanism Untreated Control ADSC IA Treated Group Leu et al, J Translational Med 2010;8(63).
Slide 28 Anti-fibrosis Mechanism Mouse Liver Fibrosis – CCl 4 – BM-MSC IV Infusions 1.Decrease TGF-B (decrease response of stellate cells) 2.Increase IL-10 (antifibrogenic cytokine) Fang et al, Transplantation 78:1;2004 Control MSC Blue = fibrosis
Slide 29 Homing and Angiogenesis Ischemia Model – Adipose Cell Therapy 7 days post ischemia - IV Laser Doppler Blood Flow Saline Control Adipose Stem Cell Treated Miranville, Circulation, 2004
Slide 30 Roles / Functions “Stem cells are injury-specific, perfectly choreographed pharmaceutical factories” Influenced by injury micro-environment “Paramedics” Dr. Arnold Caplan, Case Western Reserve University
Slide 31 ‘Activation’ of Stem Cells “In vivo use of hMSCs for therapeutic indications does not require priming of MSCs.” Annu. Rev. Pathol. Mech. Dis :457–78
Slide Goal of Regenerative Medicine is to return damaged tissue to normal state. 2. Regenerative cells function by: - Homing - Differentiation into needed tissues - Trophic stimulation of regeneration 3. Activation of stem cells is not necessary for therapeutic effects, and may be harmful. Summary - Regenerative Medicine