title: Electroporation-mediated transfer of SOX trio genes (SOX-5, SOX-6, and SOX-9) to enhance the chondrogenesis of mesenchymal stem cells.
abstract: The purpose of this study was to test the hypothesis that the SOX trio genes (SOX-5, SOX-6, and SOX-9) have a lower level of expression during the chondrogenic differentiation of mesenchymal stem cells (MSCs) compared with chondrocytes and that the electroporation-mediated gene transfer of SOX trio promotes chondrogenesis from human MSCs. An in vitro pellet culture was carried out using MSCs or chondrocytes at passage 3 and analyzed after 7 and 21 days. Then, MSCs were transfected with SOX trio genes and analyzed for the expression of chondrogenic markers after 21 days of in vitro culture. Without transforming growth factor-β1, the untransfected MSCs had a lower level of SOX trio gene and protein expression than chondrocytes. However, the level of SOX-9 gene expression increased in MSCs when treated with transforming growth factor-β1. GAG level significantly increased 7-fold in MSCs co-transfected with SOX trio, which was corroborated by Safranin-O staining. SOX trio co-transfection significantly increased COL2A1 gene and protein and decreased COL10A1 protein in MSCs. It is concluded that the SOX trio have a significantly lower expression in human MSCs than in chondrocytes and that the electroporation-mediated co-transfection of SOX trio enhances chondrogenesis and suppresses hypertrophy of human MSCs.
authors.affiliation: Department of Orthopaedics, Dongguk University Ilsan Hospital, Goyang, Republic of Korea.
authors.forename: Hye-Joung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Gun-Il
authors.initials: GI
authors.lastname: Im
mesh_terms: D000368:Aged; D017209:Apoptosis / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D020219:Chondrogenesis / Q000235:genetics*; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D005786:Gene Expression Regulation; D006025:Glycosaminoglycans / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D055747:SOX Transcription Factors / Q000235:genetics* / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D055755:SOXD Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D015415:Biomarkers; D006025:Glycosaminoglycans; D055747:SOX Transcription Factors; D055758:SOX9 Transcription Factor; D055755:SOXD Transcription Factors; D049452:Green Fluorescent Proteins; D003094:Collagen; D004247:DNA