Chondrogenic Differentiation of Human Embryonic Stem Cells by hSOX9 Overexpression = Chondrogenic Differentiation of Human Embryonic Stem Cells by hSOX9 Overexpression

다국어 초록 (Multilingual Abstract)

Previous studies have shown that the stable enhanced green fluorescence protein(EGFP) positive human embryonic stem cell(EGFP-CHA3 hESC) line was constructed using lentiviral system with the human polypeptide chain elongation factor-1alpha(EF-1α) promoter. Based this report, we inserted a hSOX9 fragment into self-inactivating( SIN) human immunodeficiency virus type-1(HIV-1)-based lentiviral vectors containing EF-1alpha promoter, and continuously induced chondrogenesis by using lentiviral vectors. It has known that hSOX9 is a transcription factor, which has high mobility group(HMG-box) that has highly homology with SRY. During chondrogenesis, hSOX9 is expressed all chondrocytes, even chondroprogenitors. After transfected virus particles into hESCs, the selected clones of transfected hESCs were aggregated and formed human embryo bodies(hEBs) for 10 days. The expression of chondrogenic specific genes such as Collagen X, Collagen II, Aggrecan, Perlecan, and Syndecan 1 were shown to highly increase by RT-PCR. Continuously, we confirmed the expression of these genes by quantitative real-time PCR. In addition, the expression of glycosaminoglycans and Collagen ll in the transfected hEBs were strongly stained by Alcian blue and immunocytochemistry. These results suggested that hSOX9-transfected hESCs were efficiently driven chondrogenesis in vitro. Taken together, the genetic modification using hSOX9 gene provides a suitable model to study basic mechanisms of differentiation into chondrogenic lineages.