title: Differentiation character of adult mesenchymal stem cells and transfection of MSCs with lentiviral vectors.
abstract: This study examined the differentiation character and pluripotency of mesenchymal stem cells (MSCs) under different conditions. Adult MSCs were initially isolated from the bone marrow of rats, cultured in vitro and identified by flow cytometry. After MSCs were transferred to osteogenic and adipogenic medium respectively, the morphological characterization of induced cells was observed. The expression of marker genes was detected by RT-PCR analysis. Then MSCs were transfected with lentiviral vectors pGC-FU-Sox9-EGFP. Enhanced green fluorescence protein (EGFP) expression and transfection efficiency were determined by fluorescence microscopy. The results demonstrated that EGFP caused no effect on the multilineage potential of adult MSCs. Sox9 gene expression of high level was maintained stable in the transfected MSCs and induced MSCs to differentiate into chondrocytes. Aggracan was positive in chondrogenic lineages and the expression of aggracan and type collagen II was significantly increased during MSCs chondrogenic differentiation. It was concluded that Sox9 gene-modified adult MSCs may be promising candidate cells for further studies on tissue engineering. EGFP facilitates the research on MSCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.
authors.affiliation: Department of Integrated Chinese and Western Medicine, Huazhong University of Science and Technology, Wuhan 430022, China. zhllyc73@163.com
authors.forename: Xiayi
authors.initials: X
authors.lastname: Zhang
authors.forename: Jinsong
authors.initials: J
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Nie
authors.forename: Ke
authors.initials: K
authors.lastname: Jiang
authors.forename: Zhikun
authors.initials: Z
authors.lastname: Zhen
authors.forename: Jianjun
authors.initials: J
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Shen
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology*; D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D024043:Collagen Type II / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D055758:SOX9 Transcription Factor / Q000235:genetics; D023822:Tissue Engineering; D014162:Transfection
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D055758:SOX9 Transcription Factor; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins