title: Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep.
abstract: OBJECTIVE
  To evaluate different protocols to isolate stem cells from ovine umbilical cord blood and adipose tissue.
  
  
  METHODS
  There were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. There were used 3 protocols for obtainment and culture of umbilical cord blood stem cells and 4 protocols for ovine adipose tissue stem cells.
  
  
  RESULTS
  It was possible to observe only one successful protocol for the obtainment of umbilical cord blood stem cells. When analyzing the techniques used to obtain adipose tissue stem cells, only one of the methods was effective as well. Through colony forming unit assay, there were obtained 58 colonies of cells after seven days in culture. Flow citometry tests revealed the cells were positive to CD44 and exhibited negative reaction to CD38, CD45, CD41/61. These cells showed a growth curve with very well defined phases LOG, LAG and PLATEAU. This phases are typically seem in mesenchymal stem cells growth curves.
  
  
  CONCLUSIONS
  The isolation and culture of mesenchymal stem cells from ovine umbilical cord blood are complex and request more detailed assays. Stem cells from fat tissue sheep showed mesenchymal characteristics, according to their cell growth curve, ability to origin colonies of fibroblastoid cells and positive reactivity with the antibody CD44 by flow citometry.
authors.affiliation: Surgery Department, Faculty of Veterinary Medicine and Animal Science, USP, Sao Paulo, Brazil.
authors.forename: Leandro
authors.initials: L
authors.lastname: Fadel
authors.forename: Brunno Rosa
authors.initials: BR
authors.lastname: Viana
authors.forename: Matheus Levi Tajra
authors.initials: ML
authors.lastname: Feitosa
authors.forename: Anna Caroline Mazeto
authors.initials: AC
authors.lastname: Ercolin
authors.forename: Kelly Cristine Santos
authors.initials: KC
authors.lastname: Roballo
authors.forename: Juliana Barbosa
authors.initials: JB
authors.lastname: Casals
authors.forename: Naira Caroline Godoy
authors.initials: NC
authors.lastname: Pieri
authors.forename: Flávio Vieira
authors.initials: FV
authors.lastname: Meirelles
authors.forename: Daniele dos Santos
authors.initials: Ddos S
authors.lastname: Martins
authors.forename: Maria Angélica
authors.initials: MA
authors.lastname: Miglino
authors.forename: Carlos Eduardo
authors.initials: CE
authors.lastname: Ambrósio
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line / Q000166:cytology; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D015203:Reproducibility of Results; D012756:Sheep; D013997:Time Factors
chemical_list: D015415:Biomarkers