外胚间充质干细胞构建组织工程骨骼肌的应用研究

Application of Construction of Tissue Engineering Skeletal Muscle by Cultured Ectomesenchymal Stem Cell

目的:探讨利用大鼠颌突外胚间充质干细胞构建组织工程骨骼肌的可行性,并观察对骨骼肌缺损的修复重建的促进效应。方法:取妊娠E 11.5胎鼠颌突外胚间充质干细胞,纯化后在含5ml/L体积浓度二甲基亚砜的DMEM/F12培养基中诱导分化为骨骼肌样细胞,将细胞种植于BAM膜上培养形成组织工程骨骼肌。将其移植入大鼠骨骼肌缺损模型,手术后14 d观察骨骼肌恢复情况,同期进行组织学及免疫组化染色鉴定。结果:经诱导后外胚间充质干细胞可向骨骼肌样细胞转化,构建的组织工程骨骼肌可加速缺损的修复重建,组织学染色显示外胚间充质干细胞具有正常骨骼肌的组织形态,可表达成肌相关蛋白MyOD。结论:诱导后的外胚间充质干细胞可作为种子细胞构建组织工程骨骼肌,本实验为临床肌肉的缺损修复奠定了理论基础。

Objective： To investigate the feasibility of constructing tissue engineering skeletal muscle with ectomesenchymal stem cells and the improvement on reconstruction of skeletal muscle defect. Methods： The ectomesenchymal stem cells were derived from E 11.5 fetal rat mandibular process, and then, these cells were differentiated into skeleton-muscle-like cells with 5 ml/L DMSO. Tissue engineering skeletal muscle constructed with BAM membrane and differentiated cells were transplanted to the site of muscle defect. After 14 days, all specimens were investigated with histological and immunohistochemical staining. Results： The ectomesenchymal stem cells can be differentiated to skeleton-muscle-like cells by DMSO and the tissue engineering skeletal muscle can accelerate the repair and reconstruction of muscle. The tissue engineered muscles were similar to normal muscle in the characteristic of histological and morphological feature and express the positive signal of MyoD after 14 day transplantation. Conclusion： Ectomesenchymal stem cells were the ideal seeding cells in construct tissue-engineering muscle. The experimental study can provide the potential theory foundation and application value for the repair of muscle defect in the clinic.