Chondrogenesis enhanced by overexpression of sox9 gene in mouse bone marrow-derived mesenchymal stem cells.
We investigated chondrogenesis of cell-mediated sox9 gene therapy as a new treatment regimen for cartilage regeneration. pIRES2-EGFP vector containing a full-length mouse sox9 cDNA was transfected into bone marrow-derived mesenchymal stem cells (MSCs) by lipofection and chondrogenic differentiation of these cells was evaluated. In vitro high density micromass culture of these sox9 transfected MSCs demonstrated that a matrix-rich micromass aggregate with EGFP expressing MSCs was positively stained by Alcian blue and type II collagen. Next, sox9 transfected MSCs were loaded into the diffusion chamber and transplanted into athymic mice to analyze in vivo chondrogenesis. A massive tissue formation in about 2mm diameter was visible in the chamber after 4 weeks transplantation. Histological examinations demonstrated that both Alcian blue and type II collagen were positively stained in the extracellular matrix of the mass while type X collagen was not stained. These results indicated that cell-mediated sox9 gene therapy could be a novel strategy for hyaline cartilage damage.
Tsuchiya
Hiroki
H
Department of Orthopaedic Surgery, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan. hiroki-t@gc4.so-net.ne.jp
Kitoh
Hiroshi
H
Sugiura
Fumiaki
F
Ishiguro
Naoki
N
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D048429:Cell Size; D017690:Cell Transplantation; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology*; D046508:Culture Techniques / Q000379:methods; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D006609:High Mobility Group Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D055758:SOX9 Transcription Factor; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D006609:High Mobility Group Proteins; D011993:Recombinant Fusion Proteins; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; D014157:Transcription Factors