Effect Of TAGLN On The Migration Capacity Of Wuzhishan Mini Pigs’ Bone Marrow Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) are pluripotent adult stem cells, which can beeasily isolated from bone marrow and then expanded in vitro. It is important for MSCsbeing used for self-repair and tissue regenerative therapy in human being. As the seedcells in the adult stem cells family, MSCs have become the focus and advancing front inthe field of recent medical and cell engineering. Bone marrow mesenchymal stem cell(BMMSC) is the most widely used MSC in the life sciences and cell engineering atpresent. The poor immunogenicity ability of BMMSCs played the pivotal role in thetissue regenerative medicine and the damage repair. They not only have great potentialability of differentiation and high plasticity, but also can synthesize and secrete a varietyof factors in the bone marrow, which can provid a good environment for the division,proliferation and differentiation of hematopoietic stem cells via the interaction betweencells. However, the study on influence of migration ability of BMMSCs, especially forthe Wuzhishan miniature pig (WZSP) BMMSCs, was very limited. For this purpose, wefurther studied the factors that influenced the migration of WZSP BMMSCs.In this study, we had studied the role of TAGLN in regulating the migration abilityof BMMSCs of WZSP in vitro. The shRNA eukaryotic expression vector for TAGLNwas constructed and transfected into BMMSCs, which was used to further study theinterference effect on the target gene TAGLN by this expression vector. It provided atheory basis for MSCs in the application of the tissue damage repairing and organreconstruction. TAGLN gene was used as the target gene and specific shRNAeukaryotic expression vector for TAGLN was constructed. This vector with endotoxinfree was transfected into BMMSCs through electroporate. And then, the expressionlevel of TAGLN in BMMSCs was detected using RT-PCR and Western blot aftertransfection. The migration capacity for the BMMSCs was estimated using the scratchassay and transwell in vitro migration model. These methods could be used to explorethe influence of knockdown TAGLN on MSCs’ migration capacity. RT-PCR assayshowed that the expression of TAGLN in BMMSCs transfected by specific shRNAeukaryotic vector for TAGLN was less than that of BMMSCs transfected by emptyvector. Western blot showed that the protein expression of TAGLN in BMMSCstransfected by specific shRNA eukaryotic vector for TAGLN was also lower than that ofBMMSCs transfected by empty vector. The scratch assay and Transwell in vitro migration model assay showed that the migration ability of BMMSCs transfected byspecific shRNA eukaryotic vector for TAGLN was lower than that of BMMSCstransfected by empty vector. These experiments proved that the TAGLN could regulateMSCs’ migration ability.There was positive correlation between TAGLN expression level in MSCs and themigration capacity of BMMSCs from WZSP and the cell line which was stableinterfered by specific shRNA eukaryotic expression vector for TAGLN was obtained. Itwas the first time that the influence of TAGLN on the migration of MSCs from WZSPwas analyzed in this study, which established the theory foundation for farther researchon effect of TAGLN on the MSCs migration. The result showed that the specific shRNAinterfered the expression of TAGLN gene in BMMSCs, which degraded the BMMSCs’migration ability from WZSP. In a word, Transgelin could regulate the BMMSCs’migration ability of WZSP, which also provid the experimental and theoretical basis forthe application of MSCs in tissue damage repair, organ grafting and so on.