title: Therapeutic potential of ixmyelocel-T, an expanded autologous multicellular therapy for treatment of ischemic cardiovascular diseases.
abstract: BACKGROUND
  Bone marrow derived cellular therapies are an emerging approach to promoting therapeutic angiogenesis in ischemic cardiovascular disease. However, the percentage of regenerative cells in bone marrow mononuclear cells (BMMNCs) is small, and large amounts of BMMNCs are required. Ixmyelocel-T, an expanded autologous multicellular therapy, is manufactured from a small sample of bone marrow aspirate. Ixmyelocel-T contains expanded populations of mesenchymal stromal cells (MSCs) and M2-like macrophages, as well as many of the CD45+ cells found in the bone marrow. It is hypothesized that this expanded multi-cellular therapy would induce angiogenesis and endothelial repair.
  
  
  METHODS
  A rat model of hind limb ischemia was used to determine the effects of ixmyelocel-T on blood flow recovery. To further determine the effects on endothelial cells, ixmyelocel-T was co-cultured with human umbilical vein endothelial cells (HUVEC) in non-contacting Transwell® inserts.
  
  
  RESULTS
  Co-culture of HUVECs with ixmyelocel-T resulted secretion of a variety of pro-angiogenic factors. HUVECs stimulated by ixmyelocel-T exhibited enhanced migration, proliferation, and branch formation. Ixmyelocel-T co-culture also resulted in increased endothelial nitric oxide synthase (eNOS) expression and nitric oxide (NO) production. In tumor necrosis factor alpha (TNFα)-stimulated HUVECs, ixmyelocel-T co-culture decreased apoptosis and reactive oxygen species generation, increased super oxide dismutase activity, and decreased nuclear factor kappa B (NFκB) activation. Treatment with ixmyelocel-T in a rat model of hind limb ischemia resulted in significantly increased blood flow perfusion and capillary density, gene expression and plasma levels of the anti-inflammatory cytokine interleukin (IL)-10, plasma nitrates, plasma platelet-derived growth factor (PDGF)-BB, vascular endothelial growth factor (VEGF) expression, and significantly decreased plasma thiobarbituric acid reactive substances (TBARS).
  
  
  CONCLUSIONS
  This work demonstrates that ixmyelocel-T interacts with endothelial cells in a paracrine manner, resulting in angiogenesis and endothelial protection. This data suggests that ixmyelocel-T could be useful for promoting of angiogenesis and tissue repair in ischemic cardiovascular diseases. In conclusion, ixmyelocel-T therapy may provide a new aspect of therapeutic angiogenesis in this patient population where expanded populations of regenerative cells might be required.
authors.affiliation: Aastrom Biosciences, Domino's Farms, Lobby K 24 Frank Lloyd Wright Drive, Ann Arbor, MI, 48105, USA. kledford@aastrom.com.
authors.forename: Kelly J
authors.initials: KJ
authors.lastname: Ledford
authors.affiliation: Aastrom Biosciences, Domino's Farms, Lobby K 24 Frank Lloyd Wright Drive, Ann Arbor, MI, 48105, USA. emurphy@aastrom.com.
authors.forename: Nikki
authors.initials: N
authors.lastname: Murphy
authors.affiliation: Aastrom Biosciences, Domino's Farms, Lobby K 24 Frank Lloyd Wright Drive, Ann Arbor, MI, 48105, USA. frankthescientist@gmail.com.
authors.forename: Frank
authors.initials: F
authors.lastname: Zeigler
authors.affiliation: Aastrom Biosciences, Domino's Farms, Lobby K 24 Frank Lloyd Wright Drive, Ann Arbor, MI, 48105, USA. ronnda@me.com.
authors.forename: Ronnda L
authors.initials: RL
authors.lastname: Bartel
authors.affiliation: Aastrom Biosciences, Domino's Farms, Lobby K 24 Frank Lloyd Wright Drive, Ann Arbor, MI, 48105, USA. rtubo@vcel.com.
authors.forename: Ross
authors.initials: R
authors.lastname: Tubo
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D000077214:Becaplermin; D001854:Bone Marrow Cells / Q000502:physiology; D016026:Bone Marrow Transplantation; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004195:Disease Models, Animal; D004789:Enzyme Activation / Q000187:drug effects; D006614:Hindlimb / Q000098:blood supply* / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000097:blood; D007511:Ischemia / Q000473:pathology / Q000628:therapy; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008264:Macrophages / Q000637:transplantation; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000502:physiology; D016328:NF-kappa B / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000502:physiology*; D009566:Nitrates / Q000097:blood; D009569:Nitric Oxide / Q000096:biosynthesis; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis; D019899:Paracrine Communication / Q000502:physiology; D020574:Proto-Oncogene Proteins c-sis / Q000096:biosynthesis / Q000097:blood; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000097:blood; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology
chemical_list: D016328:NF-kappa B; D009566:Nitrates; D020574:Proto-Oncogene Proteins c-sis; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D016753:Interleukin-10; D000077214:Becaplermin; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D013482:Superoxide Dismutase; D017493:Leukocyte Common Antigens