| Mesenchymal stem cells (MSCs) have multidirectional differentiation, migration andimmunosuppressive properties. As seed cells, they are widely used in tissue engineering, cellengineering. Bone marrow mesenchymal stem cells (BMMSCs), as one kind of MSCs, are most used.However, BMMSCs have a series of shortcomings, for example, hurting donors while gathering cells.These shortcomings severely limit the clinical application of BMMSCs. Therefore, looking for othercells which can replace BMMSCs becomes an important issue. Because of lower immune activity,umbilical cord mesenchymal stem cells (UCMSCs), as the ideal substitute of BMMSCs, have more andmore attention. Whether UCMSCs can completely replace BMMSCs is the key for whether UCMSCscan be widely used. But, study on the differences between UCMSCs and BMMSCs, especially on thedifferences between Wuzhishan minipig (WZSP) s’ BMMSCs and UCMSCs is still very limited.Therefore, identification the differential proteins of BMMSCs and UCMSCs, and then exploring therelated mechanism has great significance for the application of MSCs.In this experiment, MSCs were isolated from Wuzhishan minipig (WZSP) s’ umbilical cord(UCMSCs) and bone marrow (BMMSCs) respectively. We identified MSCs from the following parts:cell morphology, the potential of osteogenesis differentiation and adipogenesis differentiation anddetections of several surface antigens. To screen the differential proteins, we compared the proteinswhich expressed in UCMSCs and BMMSCs by iTRAQ method. After choosing one differential protein,vimentin, we explored the effect of vimentin to the migration of MSCs by RNAi, gene overexpression,transwell, qPCR and western blot techniques.The results showed that: MSCs isolated from WZSP were adherent cells. The shape was spindle orpolygon, similar to the shape of mechanocyte. The results of staining with Oil red O and alizarin red Sproved that cells which were suffered adipose and osteogenesis inducing culture were both positive after21days. Flow cytometric analysis results showed that both BMMSCs and UCMSCs expressed specificsurface antigens, for example, CD90, and none expressed surface antigens of hematopoietic stem cellsCD34and leucocyte antigen CD45. Using iTRAQ method, we screened131kinds of differentialproteins. After analysis of these proteins in retrieval, we found they were associated with a variety ofphysiological functions, such as cellular homeostasis, signal transduction, transcription regulation, cellproliferation, adhesion, differentiation and cell migration etc. Vimentin, related to cell migration, is akind of differentially expressed proteins between BMMSCs and UCMSCs. The expression amount ofvimentin in BMMSCs is higher than that in UCMSCs. By establishing in vitro transwell migrationmodel and scratches healing experiment, we found that BMMSCs have the higher migration level thanUCMSCs. The gene coding vimentin is called vim gene. After using RNAi technology to lower vim inBMMSCs, using lentivirus to overexpress vim gene in UCMSCs respectively, we detected the changeof migration of BMMSCs and UCMSCs. The results showed that the migration ability of BMMSCs inwhich vim gene was knocked down dropped, and the migration ability of UCMSCs in which vim genewas overexpressed increased. The results showed that vimentin can regulate MSCs migration ability.There were positive correlation between vimentin and WZSP MSCs migration. The study first explored the effect of vimentin on WZSP MSCs migration. It lays a foundation for further study the mechanismthat vimentin affect migration of MSCs. At the same time, it also provides experimental basis andtheoretical basis for better applicating MSCs in the tissue damage repair, reconstruction of organs andclinical treatment. |
