title: [In vitro differentiation of synovial-derived mesenchymal stem cells infected by adenovirus vector mediated by bone morphogenetic protein 2/7 genes into fibrocartilage cells in rabbits].
abstract: OBJECTIVE
  To investigate the feasibility of rabbit synovial-derived mesenchymal stem cells (SMSCs) differentiating into fibrocartilage cells by the recombinant adenovirus vector mediated by bone morphogenetic protein 2/7 (BMP-2/7) genes in vitro.
  
  
  METHODS
  SMSCs were isolated and purified from 3-month-old New Zealand white rabbits [male or female, weighing (2.1 +/- 0.3) kg]; the morphology was observed; the cells were identified with immunocytological fluorescent staining, flow cytometry, and cell cycles. The adipogenic, osteogenic, and chondrogenic differentiations were detected. The recombinant plasmid of pAdTrack-BMP-2-internal ribosome entry site (IRES)-BMP-7 was constructed and then was used to infect SMSCs. The cell DNA content and the oncogenicity were tested to determine the safety. Then infected SMSCs were cultured in incomplete chondrogenic medium in vitro. Chondrogenic differentiation of infected SMSCs was detected by RT-PCR, immunofluorescent staining, and toluidine blue staining.
  
  
  RESULTS
  SMSCs expressed surface markers of stem cells, and had multi-directional potential. The transfection efficiency of SMSCs infected by recombinant plasmid of pAdTrack-BMP-2-IRES-BMP-7 was about 70%. The safety results showed that infected SMSCs had normal double time, normal chromosome number, and normal DNA content and had no oncogenicity. At 21 days after cultured in incomplete chondrocyte medium, RT-PCR results showed SMSCs had increased expressions of collegan type I and collegan type II, particularly collegan type II; the expressions of RhoA and Sox-9 increased obviously. Immunofluorescent staining and toluidine blue staining showed differentiation of SMSCs into fibrocartilage cells.
  
  
  CONCLUSIONS
  It is safe to use pAdTrack-BMP-2-IRES-BMP-7 for infecting SMSCs. SMSCs infected by pAdTrack-BMP-2-IRES-BMP-7 can differentiate into fibrocartilage cells spontaneously in vitro.
authors.affiliation: Department of Joint Surgery, Shanghai Changzheng Hospital, Shanghai, 200003, P.R. China.
authors.forename: Peiliang
authors.initials: P
authors.lastname: Fu
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Haishan
authors.initials: H
authors.lastname: Wu
authors.forename: Ruijun
authors.initials: R
authors.lastname: Cong
authors.forename: Song
authors.initials: S
authors.lastname: Chen
authors.forename: Zheru
authors.initials: Z
authors.lastname: Ding
authors.forename: Kaimen
authors.initials: K
authors.lastname: Hu
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D005260:Female; D051445:Fibrocartilage / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013583:Synovial Membrane / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection
chemical_list: D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II