J Reconstr Microsurg 2015; 31(02): 132-138
DOI: 10.1055/s-0034-1395494
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Role of Anatomical Region and Hypoxia on Angiogenic Markers in Adipose-Derived Stromal Cells

Jacob Rinkinen
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Jeffrey Lisiecki
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Eboda Oluwatobi
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Jonathan Peterson
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Sara De La Rosa
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Kavitha Ranganathan
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Stewart C. Wang
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Paul S. Cederna
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
,
Benjamin Levi
1   Department of Surgery, Section of Plastic Surgery, University of Michigan, Ann Arbor, Michigan
› Author Affiliations
Further Information

Publication History

25 March 2014

31 July 2014

Publication Date:
12 December 2014 (online)

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.

Patients and 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.

Conclusion 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.

Note

University of Michigan Institutional Review Board approval was obtained before commencement of the study (IRB # HUM00051190).


 
  • References

  • 1 Zuk PA, Zhu M, Ashjian P , et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002; 13 (12) 4279-4295
  • 2 Gir P, Oni G, Brown SA, Mojallal A, Rohrich RJ. Human adipose stem cells: current clinical applications. Plast Reconstr Surg 2012; 129 (6) 1277-1290
  • 3 Yoshimura K, Suga H, Eto H. Adipose-derived stem/progenitor cells: roles in adipose tissue remodeling and potential use for soft tissue augmentation. Regen Med 2009; 4 (2) 265-273
  • 4 Gimble JM, Katz AJ, Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res 2007; 100 (9) 1249-1260
  • 5 Yoshimura K, Shigeura T, Matsumoto D , et al. Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates. J Cell Physiol 2006; 208 (1) 64-76
  • 6 Zuk PA, Zhu M, Mizuno H , et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7 (2) 211-228
  • 7 Caplan AI, Bruder SP. Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. Trends Mol Med 2001; 7 (6) 259-264
  • 8 Yoshimura K, Asano Y, Aoi N , et al. Progenitor-enriched adipose tissue transplantation as rescue for breast implant complications. Breast J 2010; 16 (2) 169-175
  • 9 Yoshimura K, Sato K, Aoi N, Kurita M, Hirohi T, Harii K. Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells. Aesthetic Plast Surg 2008; 32 (1) 48-55 , discussion 56–57
  • 10 Yoshimura K, Sato K, Aoi N , et al. Cell-assisted lipotransfer for facial lipoatrophy: efficacy of clinical use of adipose-derived stem cells. Dermatol Surg 2008; 34 (9) 1178-1185
  • 11 Kim M, Kim I, Lee SK, Bang SI, Lim SY. Clinical trial of autologous differentiated adipocytes from stem cells derived from human adipose tissue. Dermatol Surg 2011; 37 (6) 750-759
  • 12 Aksu AE, Rubin JP, Dudas JR, Marra KG. Role of gender and anatomical region on induction of osteogenic differentiation of human adipose-derived stem cells. Ann Plast Surg 2008; 60 (3) 306-322
  • 13 Schipper BM, Marra KG, Zhang W, Donnenberg AD, Rubin JP. Regional anatomic and age effects on cell function of human adipose-derived stem cells. Ann Plast Surg 2008; 60 (5) 538-544
  • 14 Van Harmelen V, Röhrig K, Hauner H. Comparison of proliferation and differentiation capacity of human adipocyte precursor cells from the omental and subcutaneous adipose tissue depot of obese subjects. Metabolism 2004; 53 (5) 632-637
  • 15 Tchkonia T, Giorgadze N, Pirtskhalava T , et al. Fat depot origin affects adipogenesis in primary cultured and cloned human preadipocytes. Am J Physiol Regul Integr Comp Physiol 2002; 282 (5) R1286-R1296
  • 16 Kelly IE, Han TS, Walsh K, Lean ME. Effects of a thiazolidinedione compound on body fat and fat distribution of patients with type 2 diabetes. Diabetes Care 1999; 22 (2) 288-293
  • 17 Björntorp P. Abdominal obesity and the development of noninsulin-dependent diabetes mellitus. Diabetes Metab Rev 1988; 4 (6) 615-622
  • 18 Monzon JR, Basile R, Heneghan S, Udupi V, Green A. Lipolysis in adipocytes isolated from deep and superficial subcutaneous adipose tissue. Obes Res 2002; 10 (4) 266-269
  • 19 Chung HM, Won CH, Sung JH. Responses of adipose-derived stem cells during hypoxia: enhanced skin-regenerative potential. Expert Opin Biol Ther 2009; 9 (12) 1499-1508
  • 20 Lee EY, Xia Y, Kim WS , et al. Hypoxia-enhanced wound-healing function of adipose-derived stem cells: increase in stem cell proliferation and up-regulation of VEGF and bFGF. Wound Repair Regen 2009; 17 (4) 540-547
  • 21 Kim WS, Park BS, Sung JH , et al. Wound healing effect of adipose-derived stem cells: a critical role of secretory factors on human dermal fibroblasts. J Dermatol Sci 2007; 48 (1) 15-24
  • 22 Nissen NN, Gamelli RL, Polverini PJ, DiPietro LA. Differential angiogenic and proliferative activity of surgical and burn wound fluids. J Trauma 2003; 54 (6) 1205-1210 , discussion 1211
  • 23 Planat-Benard V, Silvestre JS, Cousin B , et al. Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives. Circulation 2004; 109 (5) 656-663
  • 24 Spalding KL, Arner E, Westermark PO , et al. Dynamics of fat cell turnover in humans. Nature 2008; 453 (7196) 783-787
  • 25 Michaels V J, Rubin JP. Discussion. A comparative analysis and systematic review of the wound-healing milieu: implications for body contouring after massive weight loss. Plast Reconstr Surg 2009; 124 (5) 1683-1684
  • 26 Albino FP, Koltz PF, Gusenoff JA. A comparative analysis and systematic review of the wound-healing milieu: implications for body contouring after massive weight loss. Plast Reconstr Surg 2009; 124 (5) 1675-1682
  • 27 Shermak MA, Chang D, Magnuson TH, Schweitzer MA. An outcomes analysis of patients undergoing body contouring surgery after massive weight loss. Plast Reconstr Surg 2006; 118 (4) 1026-1031
  • 28 Levi B, Nelson ER, Hyun JS , et al. Enhancement of human adipose-derived stromal cell angiogenesis through knockdown of a BMP-2 inhibitor. Plast Reconstr Surg 2012; 129 (1) 53-66
  • 29 Kopp HG, Ramos CA, Rafii S. Contribution of endothelial progenitors and proangiogenic hematopoietic cells to vascularization of tumor and ischemic tissue. Curr Opin Hematol 2006; 13 (3) 175-181
  • 30 Cao Y, Sun Z, Liao L, Meng Y, Han Q, Zhao RC. Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun 2005; 332 (2) 370-379
  • 31 Chiarugi V, Magnelli L, Chiarugi A, Gallo O. Hypoxia induces pivotal tumor angiogenesis control factors including p53, vascular endothelial growth factor and the NFkappaB-dependent inducible nitric oxide synthase and cyclooxygenase-2. J Cancer Res Clin Oncol 1999; 125 (8-9) 525-528
  • 32 Royds JA, Dower SK, Qwarnstrom EE, Lewis CE. Response of tumour cells to hypoxia: role of p53 and NFkB. Mol Pathol 1998; 51 (2) 55-61
  • 33 Chung CW, Marra KG, Li H , et al. VEGF microsphere technology to enhance vascularization in fat grafting. Ann Plast Surg 2012; 69 (2) 213-219
  • 34 Lu F, Mizuno H, Uysal CA, Cai X, Ogawa R, Hyakusoku H. Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg 2008; 121 (1) 50-58
  • 35 de Vries C, Escobedo JA, Ueno H, Houck K, Ferrara N, Williams LT. The fms-like tyrosine kinase, a receptor for vascular endothelial growth factor. Science 1992; 255 (5047) 989-991
  • 36 Amos PJ, Mulvey CL, Seaman SA , et al. Hypoxic culture and in vivo inflammatory environments affect the assumption of pericyte characteristics by human adipose and bone marrow progenitor cells. Am J Physiol Cell Physiol 2011; 301 (6) C1378-C1388
  • 37 Shalaby F, Rossant J, Yamaguchi TP , et al. Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 1995; 376 (6535) 62-66