title: In vitro and in vivo induction of bone formation based on adeno-associated virus-mediated BMP-7 gene therapy using human adipose-derived mesenchymal stem cells.
abstract: OBJECTIVE
  To determine whether adeno-associated virus (AAV)-2-mediated, bone morphogenetic protein (BMP)-7-expressing human adipose-derived mesenchymal stem cells (ADMS) cells would induce bone formation in vitro and in vivo.
  
  
  METHODS
  ADMS cells were harvested from patients undergoing selective suction-assisted lipectomy and transduced with AAV carrying the human BMP-7 gene. Non-transduced cells and cells transduced with AAV serotype 2 carrying the enhanced green fluorescence protein gene served as controls. ADMS cells were qualitatively assessed for the production of BMP-7 and osteocalcin, and subjected to alkaline phosphatase (ALP) and Chinalizarin staining. A total of 2.5 x 10(6) cells mixed with type I collagen were implanted into the hind limb of severe combined immune-deficient (SCID) mice and subjected to a histological analysis 3 weeks post implantation.
  
  
  RESULTS
  Transfection of the ADMS cells achieved an efficiency of 99% at d 7. Transduction with AAV2-BMP-7 induced the expression of BMP-7 until d 56, which was markedly increased by d 7. The cells were positively stained for ALP. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. A histological examination demonstrated that implantation of BMP-7-expressing ADMS cells could induce new bone formation in vivo.
  
  
  CONCLUSIONS
  The present in vitro and in vivo study demonstrated that human ADMS cells would be a promising source of autologous mesenchymal stem cells for BMP gene therapy and tissue engineering.
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Kang
authors.forename: Wei-Ming
authors.initials: WM
authors.lastname: Liao
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Yuan
authors.forename: Pu-Yi
authors.initials: PY
authors.lastname: Sheng
authors.forename: Long-Juan
authors.initials: LJ
authors.lastname: Zhang
authors.forename: Xiang-Wei
authors.initials: XW
authors.lastname: Yuan
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D048430:Cell Shape; D017690:Cell Transplantation; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology*
chemical_list: D055419:Bone Morphogenetic Protein 7; D015675:Osteocalcin