title: [Green fluorescent protein as a tracer of bone marrow stromal cells in bone tissue engineering in rhesus].
abstract: OBJECTIVE
  To observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo.
  
  
  METHODS
  Ad5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation.
  
  
  RESULTS
  The rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope.
  
  
  CONCLUSIONS
  GFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.
authors.affiliation: Department of Orthopedics, 187 Central Hospital of PLA, Haikou 571159, China.
authors.forename: Qun-Li
authors.initials: QL
authors.lastname: Wang
authors.forename: Guo-Xian
authors.initials: GX
authors.lastname: Pei
authors.forename: Xiong
authors.initials: X
authors.lastname: Yun
authors.forename: Dan
authors.initials: D
authors.lastname: Jin
authors.forename: Kuan-Hai
authors.initials: KH
authors.lastname: Wei
authors.forename: Gao-Hong
authors.initials: GH
authors.lastname: Ren
mesh_terms: D000818:Animals; D018786:Bone Substitutes; D002454:Cell Differentiation; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008253:Macaca mulatta; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D018613:Microscopy, Confocal; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection
chemical_list: D018786:Bone Substitutes; D049452:Green Fluorescent Proteins