RESUMO
Objective To screen the differential expression genes of bone marrow MSCs stimulated by electromagnetic field(EMF)with osteogenesis microarray analysis,and to study the underlying mechanism that EMF promotes the differentiation of bone marrow MSCs. Methods The Sprague-Dawley rat bone marrow MSCs were isolated and cultured in vitro.The third-passage cells who were stimulated by EMF and served as the stimulated group,and those who were not stimulated by the EMF served as the controls.Total RNA was extracted and purified,then it was used to synthesize cDNA and cRNA.The eRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray,respectively.The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the obtained data were analyzed using the web-based completely integrated GEArray Expression Analysis Suite.RT-PCR was used to identify the chosen genes BMP1,VDR and EGF. Results Nineteen differential expression genes were found between the stimulated group and the control group.There were 6 upregulated and 13 downregulated genes in the stimulated group.Semi-quantitative RT-PCR confirmed that the expression levels of BMP1,VDR mRNA in the stimulated group were upregulated and EGF downregulated. Conclusion The gene expression profiles about osteogenesis of the bone marrow MSCs were changed after EMF intervention(1 5 Hz,1 mT).These genes are involved in the difierentiation of bone marrow MSCs into osteoblast.These results provide deeper insight into the mechanism that EMF exposure facilitates the in vitro differentiation of bone marrow MSCs.
RESUMO
In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells (MSCs) stimulated by electromagnetic field (EMF) with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-my film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes: Bmpl, BmpT, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expres- sions of Bmpl, Bmp7 mRNA of the stimulated group were up-regulated (P<0.05) and those of Egf, Egfr were down-regulated (P<0.05). It was suggested that the gene expression profiles of osteogene- sis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell ad- hesion etc.
