title: Role of anatomical region and hypoxia on angiogenic markers in adipose-derived stromal cells.
abstract: BACKGROUND
  Recent research into adipose-derived stem cells (ASCs) suggests that anatomical location has a major impact on the metabolic profile and differentiation capacity of ASCs. By having a better understanding of how various ASCs respond to cellular stressors such as hypoxia, which are induced during routine surgical procedures, we can facilitate future development of cell-based therapies to improve wound healing.
  
  
  METHODS
  Human ASCs were isolated from the superficial and deep adipose layers of four patients undergoing elective abdominoplasty. ASCs were cultured in hypoxic (1% O2, 5% CO2, and 94% N2) conditions. After 12 and 48 hours, ASCs were assessed for markers of angiogenesis by mRNA levels of vascular endothelial growth factor A (VEGF-A), vascular endothelial growth factor B (VEGF-B), and hypoxia inducible factor 1 α (HIF-1α). Western blot analysis was performed to assess levels of VEGF-A, p-NF-κB, and NF-κB. In addition, in vitro analysis of angiogenesis was performed using Matrigel assay (BD Biosciences, Franklin Lakes, NJ).
  
  
  RESULTS
  We observed significant increases in deep ASC's VEGF-A, VEGF-B, and HIF-1α mRNA expression compared with the superficial layer after 24-hour hypoxia (p < 0.05). Similar results were found when examining protein expression levels, with the deep ASCs expressing significantly larger amounts of VEGF-A and p-NF-κB (p < 0.05) compared with the superficial layer.
  
  
  CONCLUSIONS
  Our results suggest that significant variations exist in the angiogenic profile of superficial and deep ASCs. We demonstrate that superficial ASCs are less prone to transcribe potent chemokines for angiogenesis, such as VEGF-A, VEGF-B, and HIF-1α and are less likely to translate VEGF-A and NF-κB. This may help with the selection of specific stem cell donor sites in future models for stem cell therapy.
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Jacob
authors.initials: J
authors.lastname: Rinkinen
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Lisiecki
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Eboda
authors.initials: E
authors.lastname: Oluwatobi
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Peterson
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Sara
authors.initials: S
authors.lastname: De La Rosa
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Kavitha
authors.initials: K
authors.lastname: Ranganathan
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Stewart C
authors.initials: SC
authors.lastname: Wang
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Cederna
authors.affiliation: Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Levi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D002454:Cell Differentiation / Q000502:physiology; D005260:Female; D006801:Humans; D000860:Hypoxia; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000032:analysis; D059630:Mesenchymal Stem Cells; D008875:Middle Aged; D016328:NF-kappa B / Q000032:analysis; D018919:Neovascularization, Physiologic; D060888:Real-Time Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis; D042561:Vascular Endothelial Growth Factor B / Q000032:analysis
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016328:NF-kappa B; D042461:Vascular Endothelial Growth Factor A; D042561:Vascular Endothelial Growth Factor B