title: Isolation, culture and detection of mesenchymal stem cells in tissues from critical limb ischemia model in rat.
abstract: BACKGROUND
  Rat mesenchymal stem cells (MSCs) represent a small portion of the cells in the stromal compartment of bone marrow and have the potential to differentiate into new blood vessel and other tissues. MSCs transplantation in tissues from critical limb ischemia model in rat may represent a therapeutic applications of vascular regeneration.
  
  
  OBJECTIVE
  The aim of this study was to isolate with a simple method the rat bone marrow stromal cells. Then the adherent cells were labeled with 5-bromo-2-deoxyuridine (BrdU, Sigma) and injected in the gastrocnemius and adductor muscle of ischemic hind limbs in order to demonstrate their presence in the critical limb ischemia model in rat.
  
  
  METHODS
  MSCs were isolated from Wistar rats, 8 weeks of age. The MSCs were labeled in vitro for later identification by adding 10 μg/mL 5-bromo-2 -deoxyuridine (BrdU, Sigma).
  
  
  RESULTS
  Small colonies of fibroblast-like cells were seen after several days of primary culture. These colonies increased in size and were subcultured after 15-18 days.
  
  
  CONCLUSIONS
  The MSCs obtained in this study presented a stable undifferentiated phenotype under normal culture conditions. MSCs are easy to isolate, culture, and detect in in vivo culture. These cells are characterized by high plasticity and could have an important role in angiogenesis.
authors.affiliation: Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania 2 Surgical Clinic, Cluj-Napoca, Romania.
authors.forename: Nicolae-Ovidiu
authors.initials: NO
authors.lastname: Grad
authors.affiliation: Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania.
authors.forename: Pall
authors.initials: P
authors.lastname: Emoke
authors.affiliation: Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania 2 Surgical Clinic, Cluj-Napoca, Romania.
authors.forename: Ion-Aurel
authors.initials: IA
authors.lastname: Mironiuc
keywords: angiogenesis; critical limb ischemia model in rat; mesenchymal stem cells