title: Mesenchymoangioblast-derived mesenchymal stromal cells inhibit cell damage, tissue damage and improve peripheral blood flow following hindlimb ischemic injury in mice.
abstract: OBJECTIVE
  Existing treatments have limited success in modifying the course of peripheral artery disease, which may eventually lead to limb-threatening ulcers and amputation. Cellular therapies have the potential to provide a new treatment option for this condition, but isolation of cells by conventional means has limitations with respect to reproducibility and scalability.
  
  
  METHODS
  Induced pluripotent stem cells (iPSCs) were differentiated into precursor cells known as mesenchymoangioblasts (MCAs) and subsequently into mesenchymal stromal cells (MSCs). Hindlimb ischemia in mice was created by ligating both the iliac and femoral arteries of one hindlimb. Immediately after surgery, each animal received intramuscular injections of 5 × 10(6) cells or media in the ischemic limb. Toe necrosis was assessed visually, and hindlimb blood flow was measured by laser Doppler using a set region of interest (ROI) and by tracing the entire foot. Myofiber heterogeneity, nuclear centralization, fatty degeneration, fibrosis and capillary angiogenesis in the gastrocnemius muscle were assessed histologically.
  
  
  RESULTS
  Blood flow in the MCA-derived MSC-treated animals was higher at each day (P <0.006), and these mice recovered faster than control animals (3.6 vs. 2.5 for set ROI; 7.5 vs. 4.1 foot tracing; slope; P <0.001). There was significantly less myofiber heterogeneity, nuclear centralization, fatty degeneration and fibrosis in MCA-derived MSC-treated animals, indicating less tissue damage.
  
  
  CONCLUSIONS
  MCA-derived MSCs improved limb blood flow, reduced necrosis and maintained muscle mass and gross muscle appearance. We conclude that MCA-derived MSCs have a significant and protective effect against ischemic insults.
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Jill M
authors.initials: JM
authors.lastname: Koch
authors.affiliation: Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Saritha S
authors.initials: SS
authors.lastname: D'Souza
authors.affiliation: Research Animal Resources Center, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Denise J
authors.initials: DJ
authors.lastname: Schwahn
authors.affiliation: Cynata Therapeutics Limited, Armadale, Victoria, Australia.
authors.forename: Ian
authors.initials: I
authors.lastname: Dixon
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA. Electronic address: th2@medicine.wisc.edu.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Hacker
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D005263:Femoral Artery / Q000254:growth & development / Q000473:pathology; D006614:Hindlimb / Q000098:blood supply / Q000293:injuries; D007083:Iliac Artery / Q000254:growth & development / Q000473:pathology; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D007511:Ischemia / Q000628:therapy*; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000293:injuries; D009336:Necrosis / Q000473:pathology; D018919:Neovascularization, Physiologic / Q000502:physiology*; D058729:Peripheral Arterial Disease / Q000628:therapy*; D012039:Regional Blood Flow / Q000502:physiology*; D015203:Reproducibility of Results
keywords: critical limb ischemia; hindlimb ischemia; induced pluripotent stem cells; mesenchymal stem cells; peripheral arterial disease; stem cell therapy