Analysis Of Human Umbilical Cord Blood Mesenchymal Stem Cells Derived Schwann-like Cells In The Gene Expression

Background :Peripheral nerve defect is common in the Department of orthopedics of trauma,the main use of nerve autograft repair for large defect,because of its own limitations can not be widely carried out in clinical.With the development of tissue engineering,by artificial nerve repair the large segment of peripheral nerve defects is expect to the best treatment.Seed cells are the core of the artificial neural research,by mesenchymal stem cells(MSCs)differentiating into Schwann-like cells relative autologous and allogeneic Schwann cells(SCs)has superiority.It’s become better way to obtain Schwann cells.MSCs have a wide range of sources,such as bone marrow,umbilical cord,umbilical cord blood,adipose tissue,because Human umbilical cord blood mesenchymal stem cells(HUCBMSCs)have their advantages more attractive to researchers.MSCs differentiating into Schwann-like cells processes is through that the regulation of gene expression induced by objective.Through the study of induced genes in the process of expression characteristics or understand the differences between them,for later further in-depth study to provide some experimental theory.Objective:Collection of HUCBMSCs,SCs and HUCBMSCs differentiating into Schwann-like cells,extraction of three kinds of cells m RNA.Screening target genes at the transcriptional level by gene chip technology,the induction of Schwann-like cells is nearer Schwann cells.Methods:According to our group before,extraction and purification of HUCBMSCs by using erythrocyte sedimentation and density gradient centrifugation,use the low enzyme digestion and differential adhesion,medium condition extraction and purification of human SCs.The identification and purity of the cells were observed by cell morphological observation and flow cytometry and meet the requirements to the next experiment,the third generation of stem cells taken before our group to optimize the use of induced artificially induced in vitro methods.Total m RNA was extracted and use Gene chip technology to analyze the gene expression.Firstly,the differential expression genes of HUCBMSCs and SCs were screened out.Comparison of these differentially expressed genes and SCs expression genes to screen out the expression of no difference between the two group,The significance of the repair of peripheral nerve injury was found in these genes and the up-regulated expression genes were found.And select some genes for RT-PCR validation.Results:1)The number of differentially expressed genes in HUCBMSCs and SCs was6331,up to 3569 genes were up-regulated and 2762 genes were down-regulated.These differentially expressed genes are induced by the factors except for cell differentiation and other factors.2)GO enrichment analysis of differentially expressed genes,10.4%,21.2%,68.4% in the cell component,molecular function and biological process.the significant enrichment of the articles was mainly concentrated in the G protein coupled receptor activity and other 27 items.3)By matching the differentially expressed genes with SCs expression,there were no differentially expressed genes in 2265,which showed Schwann-like cells more close to SCs in the level of gene expression.4)By combining the results of the above genes with the relevant literature,6 genes were identified,which were found to be significant for the repair of nerve injury,such as S100 calcium binding proteinβ,glial cell derived neurotrophic factor,early growth response 2,vinculin,protein kinase C-alpha,protein kinase C-eta.5)In the 6 genes selection of 2 were used RT-PCR technology validation,and the results with the chip test.Conclusion:1)This experiment further verified our group can be extracted prinary HUCBMSCs and SCs.2)From the perspective of the results of this experiment,HUCBMSCs differentiating into Schwann-like cells in gene expression at the level of transcription is closer to SCs.3)The 6 expression genes were selected in this experiment which are the high express of Schwann-like cells and also express of SCs,and it also has a certain significance to peripheral nerve injury repair.These genes further showed that Schwann-like cells and SCs in the gene level has certain similarities,it’s also provide a theoretical basis for further improvement.