Modification of human bone marrow mesenchymal stem cells by using cadherin-11 and core binding factor alpha-1 gene / 中国组织工程研究

ABSTRACT

BACKGROUND:Cadherin-11 which was used to modify bone marrow mesenchymal stem cells aimed to promote cell adhesion ability;while,core binding factor α1 (cbfα1) which was used to modify bone marrow mesenchymal stem cells aimed to realize ossification of multipotential stem cells and play a signalization role in cascade effect.OBJECTIVE:To construct cadherin-11 and cbfα1 gene adenovirus expression vectors so as to modify bone marrow mesenchymal stem cells,and observe the expressions of the two genes.DESIGN,TIME AND SETTING:Cell-genetics experiment was performed at Central Laboratory of Zhujiang Hospital in August 2008.MATERIALSBone marrow mesenchymal stem cells were extracted from a male patient with thoracic vertebral fracture re from Zhujiang Hospital.The patient provided the informed consent.Full length cadherin-11 cDNA plasmid was provided by Professor Takeichi,Riken Molecular Organism Center,Japan;full length cbfα1 gene plasmid was provided by Professor Ducy,Baylor Medical College,USA;pAdeasy adenovirus system was provided by Professor Bert,McKuslck-Nathans Institute of Genetic Medicine,USA.METHODS:Human bone marrow mesenchymal stem cells were separated using Percoll density gradient centrifugation combined with adherence method.Cadherin-11 and cbfα1 genes were obtained by PCR,and then they were inserted into shuttle vector of adenovirus;thereafter,the recombinant adenovirus plasmids were gained by homologous recombination.Finally,the two plasmids were re-packed to obtain recombinant adenovirus venom,and cadherin-11 and cbfα1 genes were transfected in bone marrow mesenchymal stem cells by adenovirus.MAIN OUTCOME MEASURES:The expressions of cadherin-11 and cbfα1 genes were detected by Western biotting.RESULTS:The adenovirus venom carrying the cadherin-11 and cbfα1 gene was successfully obtained.Western blotting showed that the expressions of the cadherin-11 and cbfα1 genes in bone marrow mesenchymal stem cells were remarkably increased by infection viral.CONCLUSION:The bone marrow mesenchymal stem cells may express high level of cedherin-11 and cbfα1 by gene modification of adenovirus.