TSP-1 Modified BMSCs Promote Functional Recovery In Rats With Spinal Cord Injury

Objectives:TSP-1 modified BMSCs will be made by constructing adenovirus with TSP-1 gene and transfecting BMSCs.The effects of TSP-1 modified BMSCs in promoting functional recovery in rats with spinal cord injury and the potential mechanism will be explored.Methods:The target gene fragments were acquired by compounding the array and double enzyme digestion,identified by PCR and gene sequencing.The preparation of adenovirus with TSP-1 was operated in 293 cells line.TSP-1 modified BMSCs was made by adenovirus with TSP-1 transfecting BMSCs,verified the transfection efficiency and the expression of TSP-1 in that by immunofluorescence and western blotting.The expression of GAP-43 and NF and the percentage of VSC4.1 with neurites were measured by western blotting and immunofluorescence after the post-OGD VSC4.1 co-cultured withmedium,BMSCs,and BMSCs+TSP-1.The situation of injury and later recovery was observed by BBB score measured at the 1st and 28th day after the injury and cell transplantation.Results:In the adenovirus construction and identification section,the results of compounding and double enzyme digestion demonstrated that the target gene fragments did not have any mistake.The efficiency of adenovirus transfection achieved 80%by immunofluorescence results.The expression of TSP-1 was significantly improved in TSP-1 gene modified BMSCs by western blotting.The expression of GAP-43 and NF and the percentage of VSC4.1 with neurites were obviously increased in OGD + BMSCs+ TSP-1 group by immunofluorescence and western blotting.The locomotor function of rats in SCI+BMSCs+TSP-1 group was clearly ameliorative at the 28th day.Conclusion:TSP-1 gene modified BMSCs may accelerate the neurites repair and regeneration of post-OGD VSC4.1 and contribute to the functional restoration in the SCI rats.