title: Gene Transfer and Gene Silencing in Stem Cells to Promote Chondrogenesis.
abstract: In stem cell-based chondrogenesis for articular cartilage regeneration, TGF-β3 is dosed to the stem cells to drive differentiation into chondrocytic cells. Meanwhile, type I collagen, which is endogenously expressed in some stem cells (e.g., synovium-derived mesenchymal stem cells) and upregulated by TGF-β3, poses a threat to chondrogenesis, as type I collagen may alter the components and stiffness of articular cartilage. Therefore, a wiser strategy would be to feed the cells with TGF-β3 while at the same time silencing the expression of type I collagen. In this chapter, methods for construction of adenoviral vectors and lentiviral vectors having both of the above functions are given. Their transduction into synovium-derived mesenchymal stem cells for articular cartilage engineering and following characterizations are also described.
authors.affiliation: Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Singapore.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Singapore. DAWang@ntu.edu.sg.
authors.forename: Dong-An
authors.initials: DA
authors.lastname: Wang
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D020219:Chondrogenesis / Q000235:genetics*; D024042:Collagen Type I / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D034622:RNA Interference; D044968:Regenerative Medicine / Q000379:methods*; D013583:Synovial Membrane / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D014161:Transduction, Genetic; D014162:Transfection / Q000379:methods*; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism
chemical_list: D024042:Collagen Type I; C509044:TGFB3 protein, human; D053782:Transforming Growth Factor beta3
keywords: Adenovirus; Chondrogenesis; Lentivirus; TGF-β3; Type I collagen