title: Depot-specific variation in the osteogenic and adipogenic potential of human adipose-derived stromal cells.
abstract: BACKGROUND
  Adipose-derived stromal cells hold promise for use in tissue regeneration. However, multiple facets of their biology remain unclear. The authors examined the variations in osteogenesis and adipogenesis in adipose-derived stromal cells between subcutaneous fat depots and potential molecular causes.
  
  
  METHODS
  Adipose-derived stromal cells were isolated from human patients from subcutaneous fat depots, including arm, flank, thigh, and abdomen (n = 5 patients). Osteogenic and adipogenic differentiation was performed (alkaline phosphatase, alizarin red, and oil red O staining, and quantitative real-time polymerase chain reaction). Co-cultures were established to assess the paracrine effect of human adipose-derived stromal cells on mouse osteoblasts. Finally, HOX gene expression was analyzed by quantitative real-time polymerase chain reaction.
  
  
  RESULTS
  Subcutaneous fat depots retain markedly different osteogenic and adipogenic potentials. Osteogenesis was most robust in adipose-derived stromal cells from the flank and thigh, as compared with those from the arm and abdomen (p < 0.05 by all markers examined). This was accompanied by elevations of BMP4 and BMPR1B (p < 0.05 by all markers examined). The osteogenic advantage of cells from the flank and thigh was again observed when analyzing the paracrine effects of these cells. Conversely, those cells isolated from the flank had a lesser ability to undergo adipogenic differentiation. Adipose-associated HOX genes were less expressed in flank-derived adipose-derived stromal cells.
  
  
  CONCLUSIONS
  Variations exist between fat depots in terms of adipose-derived stromal cell osteogenic and adipogenic differentiation. Differences in HOX expression and bone morphogenetic protein signaling may underlie these observations. This study indicates that the choice of fat depot derivation of adipose-derived stromal cells may be an important one for future efforts in tissue engineering.
authors.affiliation: Stanford and Los Angeles, Calif. From the Hagey Pediatric Regenerative Medicine Research Laboratory, Department of Surgery, Plastic and Reconstructive Surgery Division, Stanford University School of Medicine, and the Division of Plastic and Reconstructive Surgery, University of California, Los Angeles.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Levi
authors.forename: Aaron W
authors.initials: AW
authors.lastname: James
authors.forename: Jason P
authors.initials: JP
authors.lastname: Glotzbach
authors.forename: Derrick C
authors.initials: DC
authors.lastname: Wan
authors.forename: George W
authors.initials: GW
authors.lastname: Commons
authors.forename: Michael T
authors.initials: MT
authors.lastname: Longaker
mesh_terms: D050156:Adipogenesis; D000328:Adult; D000818:Animals; D002454:Cell Differentiation; D006801:Humans; D051379:Mice; D008875:Middle Aged; D010012:Osteogenesis; D017154:Stromal Cells / Q000166:cytology*; D050151:Subcutaneous Fat / Q000166:cytology*
references.citation: Nacamuli RP, Longaker MT. Bone induction in craniofacial defects. Orthod Craniofac Res. 2005;8:259–266.
references.citation: Kwan MD, Slater BJ, Wan DC, Longaker MT. Cell-based therapies for skeletal regenerative medicine. Hum Mol Genet. 2008;17:R93–R98.
references.citation: Eremia S, Newman N. Long-term follow-up after autologous fat grafting: Analysis of results from 116 patients followed at least 12 months after receiving the last of a minimum of two treatments. Dermatol Surg. 2000;26:1150–1158.
references.citation: Mojallal A, Shipkov C, Braye F, Breton P, Foyatier JL. Influence of the recipient site on the outcomes of fat grafting in facial reconstructive surgery. Plast Reconstr Surg. 2009;124:471–483.
references.citation: Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002;13:4279–4295.
references.citation: Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: Implications for cell-based therapies. Tissue Eng. 2001;7:211–228.
references.citation: Levi B, James AW, Wan D, Glotzbach JP, Commons GW, Longaker MT. Regulation of human adipose-derived stromal cell osteogenic differentiation by insulin-like growth factor-1 and platelet-derived growth factor-α. Plast Reconstr Surg. 2010;126:41–52.
references.citation: Malladi P, Xu Y, Yang GP, Longaker MT. Functions of vitamin D, retinoic acid, and dexamethasone on mouse adipose-derived mesenchymal cells (AMCs). Tissue Eng. 2006;12:2031–2040.
references.citation: Wan DC, Siedhoff MT, Kwan MD, Nacamuli RP, Wu BM, Longaker MT. Refining retinoic acid stimulation for osteogenic differentiation of murine adipose-derived adult stromal cells. Tissue Eng. 2007;13:1623–1631.
references.citation: Dragoo JL, Choi JY, Lieberman JR, et al. Bone induction by BMP-2 transduced stem cells derived from human fat. J Orthop Res. 2003;21:622–629.
references.citation: Wan DC, Shi YY, Nacamuli RP, Quarto N, Lyons KM, Longaker MT. Osteogenic differentiation of mouse adipose-derived adult stromal cells requires retinoic acid and bone morphogenetic protein receptor type IB signaling. Proc Natl Acad Sci USA. 2006;103:12335–12340.
references.citation: Skillington J, Choy L, Derynck R. Bone morphogenetic protein and retinoic acid signaling cooperate to induce osteoblast differentiation of preadipocytes. J Cell Biol. 2002;159:135–146.
references.citation: Knippenberg M, Helder MN, Zandieh Doulabi B, Wuisman PI, Klein-Nulend J. Osteogenesis versus chondrogenesis by BMP-2 and BMP-7 in adipose stem cells. Biochem Biophys Res Commun. 2006;342:902–908.
references.citation: Levi B, James AW, Xu Y, et al. Divergent modulation of adipose-derived stromal cell differentiation by TGF-β1 based on species of derivation. Plast Reconstr Surg. 2010;126:412–425.
references.citation: Mehlhorn AT, Niemeyer P, Kaschte K, et al. Differential effects of BMP-2 and TGF-beta1 on chondrogenic differentiation of adipose derived stem cells. Cell Prolif. 2007;40:809–823.
references.citation: Levi B, James AW, Nelson ER, et al. Human adipose derived stromal cells heal critical size mouse calvarial defects. PLoS One 2010;5:e11177.
references.citation: Cowan CM, Shi YY, Aalami OO, et al. Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol. 2004;22:560–567.
references.citation: Tchoukalova YD, Koutsari C, Karpyak MV, Votruba SB, Wendland E, Jensen MD. Subcutaneous adipocyte size and body fat distribution. Am J Clin Nutr. 2008;87:56–63.
references.citation: Tchoukalova YD, Nathanielsz PW, Conover CA, Smith SR, Ravussin E. Regional variation in adipogenesis and IGF regulatory proteins in the fetal baboon. Biochem Biophys Res Commun. 2009;380:679–683.
references.citation: Perrini S, Laviola L, Cignarelli A, et al. Fat depot-related differences in gene expression, adiponectin secretion, and insulin action and signalling in human adipocytes differentiated in vitro from precursor stromal cells. Diabetologia 2008;51:155–164.
references.citation: van Beek EA, Bakker AH, Kruyt PM, Hofker MH, Saris WH, Keijer J. Intra- and interindividual variation in gene expression in human adipose tissue. Pflugers Arch. 2007;453:851–861.
references.citation: Tchkonia T, Lenburg M, Thomou T, et al. Identification of depot-specific human fat cell progenitors through distinct expression profiles and developmental gene patterns. Am J Physiol. 2007;292:E298–E307.
references.citation: Ray H, Pinteur C, Frering V, Beylot M, Large V. Depot-specific differences in perilipin and hormone-sensitive lipase expression in lean and obese women. Lipids Health Dis. 2009;8:58.
references.citation: Palou M, Priego T, Sanchez J, Rodriguez AM, Palou A, Pico C. Gene expression patterns in visceral and subcutaneous adipose depots in rats are linked to their morphologic features. Cell Physiol Biochem. 2009;24:547–556.
references.citation: Andersson T, Söderström I, Simonyté K, Olsson T. Estrogen reduces 11beta-hydroxysteroid dehydrogenase type 1 in liver and visceral, but not subcutaneous, adipose tissue in rats. Obesity (Silver Spring) 2010;18:470–475.
references.citation: Joe AW, Yi L, Even Y, Vogl AW, Rossi FM. Depot-specific differences in adipogenic progenitor abundance and proliferative response to high-fat diet. Stem Cells 2009;27:2563–2570.
references.citation: Klöting N, Berthold S, Kovacs P, et al. MicroRNA expression in human omental and subcutaneous adipose tissue. PLoS One 2009;4:e4699.
references.citation: Festuccia WT, Blanchard PG, Turcotte V, et al. Depot-specific effects of the PPARgamma agonist rosiglitazone on adipose tissue glucose uptake and metabolism. J Lipid Res. 2009;50:1185–1194.
references.citation: 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;60:538–544.
references.citation: 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;60:306–322.
references.citation: Samulin J, Lien S, Grindflek E, Burget I, Ruyter B, Sundvold H. Depot specific differences during adipogenesis of porcine stromal-vascular cells. Cell Biol Int. 2008;32:525–531.
references.citation: 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:R1286–R1296.
references.citation: Xu Y, Malladi P, Zhou D, Longaker MT. Molecular and cellular characterization of mouse calvarial osteoblasts derived from neural crest and paraxial mesoderm. Plast Reconstr Surg. 2007;120:1783–1795.
references.citation: James AW, Xu Y, Wang R, Longaker MT. Proliferation, osteogenic differentiation, and fgf-2 modulation of posterofrontal/sagittal suture-derived mesenchymal cells in vitro. Plast Reconstr Surg. 2008;122:53–63.
references.citation: Pei L, Tontonoz P. Fat's loss is bone's gain. J Clin Invest. 2004;113:805–806.
references.citation: Lee J, Gupta DM, Pannetta NJ, et al. Elucidating mechanisms of osteogenesis in human adipose-derived stromal cells via microarray analysis. J Craniofac Surg. 2010;21:1136–1141.
references.citation: Cowan CM, Aalami OO, Shi YY, et al. Bone morphogenetic protein 2 and retinoic acid accelerate in vivo bone formation, osteoclast recruitment, and bone turnover. Tissue Eng. 2005;11:645–658.
references.citation: Dragoo JL, Lieberman JR, Lee RS, et al. Tissue-engineered bone from BMP-2-transduced stem cells derived from human fat. Plast Reconstr Surg. 2005;115:1665–1673.
references.citation: Valeri CR, Pivacek LE. Effects of the temperature, the duration of frozen storage, and the freezing container on in vitro measurements in human peripheral blood mononuclear cells. Transfusion. 1996;36:303–308.
references.citation: Thirumala S, Gimble JM, Devireddy RV. Cryopreservation of stromal vascular fraction of adipose tissue in a serum-free freezing medium. J Tissue Eng Regen Med. 2010;4:224–232.
references.citation: Guilak F, Lott KE, Awad HA, et al. Clonal analysis of the differentiation potential of human adipose-derived adult stem cells. J Cell Physiol. 2006;206:229–237.
references.citation: Gronthos S, Franklin DM, Leddy HA, Robey PG, Storms RW, Gimble JM. Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol. 2001;189:54–63.
references.citation: Katz AJ, Tholpady A, Tholpady SS, Shang H, Ogle RC. Cell surface and transcriptional characterization of human adipose-derived adherent stromal (hADAS) cells. Stem Cells. 2005;23:412–423.
references.citation: Hattori H, Sato M, Masuoka K, et al. Osteogenic potential of human adipose tissue-derived stromal cells as an alternative stem cell source. Cells Tissues Organs. 2004;178:2–12.