Experimental Study On Inducing Pre-fibroblast To Differentiate Into Muscle Cells By Transfection Of Myogenic Gene MyoD

The weight of muscles is about 40% of that of a human body. Muscle injuries are a challenging problem in traumatology, and the most frequent occurrence in the excision of tumors. Regeneration of muscles after injury has always been a heat point. Although the injury is capable of healing to some extent, an incomplete functional recovery often occurs, depending on the severity of trauma. The muscle function reduced is caused mainly by the formation of scar during healing.For decades, grafts of muscles of the same body surgically have often been used for the restoration of the injured parts, and the limit and inevitable function loss of the donor is obvious. It is necessary to look for a new way to the treatment of muscle injuries and a new material for he muscle restoration.Tissue engineering and gene therapy turn over a new leaf for the treatment and restoration of muscles after injury. It is said that muscle is anattractive target for gene therapy. Recently MyoD was found to play a key role in the early stages of myogenesis, cell differentiation, and regeneration after muscle injury. We notice that pre-fibroblasts are the most restorative cells in the granulated tissue formed after muscle injury. If the restorative fibroblasts in granulated tissue could be transformed into myogenic cells, then muscle would be repaired itself instead of scar formation, and its function could be improved. A new approach to harvest seeding cells economically is sure to be found and the muscle morphological restoration and its functional reconstruction will be solved.In this study, we used gene cloning technology to construct the eukaryotic expression vector pcDNA3/MyoD in which mouse MyoD cDNA was inserted. Mouse NIH3T3 cells were transfected with exogenetic MyoD cDNA via lipofectamine, and cloned by G418. Expression and its biological functions of mouse MyoD were testified by molecular and immunohistological methods. Myotube were found from cultures incubated a week in differentiation medium. Ecoptic myogenesis was found when the transfected cells were seeded on the back of nude mouse. A new biological membrane used as ECM and scaffold was evaluated and the cells adhered and grew well. The membrane may be useful in the muscle tissue engineering.To sum up, fibroblasts can be tansfected by MyoD cDNA into myogenic cells, which are promising in the therapy of muscle injuries.Gene therapy and tissue engineering promise to revolutionize muscle surgery.