title: In vitro and in vivo induction of bone formation based on ex vivo gene therapy using rat adipose-derived adult stem cells expressing BMP-7.
abstract: BACKGROUND
  Adipose-derived adult stem (ADAS) cells are multipotent cells capable of differentiating into osteoblasts, adipocytes and chondrocytes. The aim of this study was to determine whether BMP-7-expressing ADAS cells would elicit bone formation invitro and in vivo.
  
  
  METHODS
  ADAS cells were harvested from Lewis rats and transduced with adenovirus carrying the recombinant human bone morphogenetic protein-7 (Ad-BMP-7) gene. Untransduced cells and cells transduced with adenovirus carrying the enhanced green fluorescence protein (Ad-EGFP) gene served as controls. BMP-7 expression was assessed by RT-PCR, immunofluorescence on day 1, and Western blot on days 4, 8 and 12. Alkaline phosphatase (ALP) activity was assayed on days 2, 4, 6, 8, 10 and 12. Osteocalcin production and bone nodule formation were detected by immunohistochemistry and von Kossa stain on day 12. A total of 1 x 10(6) cells mixed with type I collagen were implanted into the subcutaneous pocket in Lewis rat and subjected to histologic analysis 1, 2 and 4 weeks post-implantation.
  
  
  RESULTS
  The Ad-BMP-7-transduced ADAS cells expressed BMP-7 at both mRNA and protein levels. ALP activity was detected in Ad-BMP-7-transduced cells from day 2 to day 12, peaking on day 8. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. Histologic examination revealed that implantation of BMP-7-expressing ADAS cells could induce new bone formation in vivo.
  
  
  CONCLUSIONS
  ADAS cells would be a promising source of adult autologous stem cells for BMP gene therapy and tissue engineering.
authors.affiliation: Department of Orthopedics, Peking University Third Hospital, Beijing, China.
authors.forename: M
authors.initials: M
authors.lastname: Yang
authors.forename: Q J
authors.initials: QJ
authors.lastname: Ma
authors.forename: G T
authors.initials: GT
authors.lastname: Dang
authors.forename: K t
authors.initials: Kt
authors.lastname: Ma
authors.forename: P
authors.initials: P
authors.lastname: Chen
authors.forename: C Y
authors.initials: CY
authors.lastname: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D000818:Animals; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis; D051381:Rats; D011917:Rats, Inbred Lew; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000096:biosynthesis*
chemical_list: C525819:BMP7 protein, human; C525821:Bmp7 protein, rat; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta; D003094:Collagen