Transplantation of Myocyte Precursors Derived from Embryonic Stem Cells Transfected with IGFII Gene in a Mouse Model of Muscle Injury
Transplantation of Myocyte Precursors Derived from Embryonic Stem Cells Transfected with IGFII Gene in a Mouse Model of Muscle Injury
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DOI：
10.1097/01.tp.0000229388.97549.55
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发表时间：
2006-08
期刊：
影响因子：
6.2
通讯作者：
H. Kamochi;M. Kurokawa;H. Yoshikawa;Yuji Ueda;C. Masuda;E. Takada;Kenji Watanabe;M. Sakakibara;Yasunori Natuki;K. Kimura;M. Beppu;H. Aoki;N. Suzuki
中科院分区：
文献类型：
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作者：
H. Kamochi;M. Kurokawa;H. Yoshikawa;Yuji Ueda;C. Masuda;E. Takada;Kenji Watanabe;M. Sakakibara;Yasunori Natuki;K. Kimura;M. Beppu;H. Aoki;N. Suzuki
背景骨骼肌组织的重建由于缺乏组织的功能替代物而受到阻碍。方法.用胰岛素样生长因子（IGF）II基因转染胚胎干（ES）细胞，并用G418筛选。分析所得细胞克隆在体外和体内的成肌分化。结果细胞表达早期和晚期肌分化标志物，包括myoD，肌细胞生成素和肌营养不良蛋白在体外。它们在细胞内磷酸化Akt，表明它们被分泌的IGFII激活。与移植未分化ES细胞的损伤小鼠和仅给予载体的损伤小鼠相比，将细胞移植到损伤的小鼠胫前肌显著改善了它们的运动功能。转染的细胞适应损伤的肌肉，形成肌营养不良蛋白阳性和阴性的MyoD和肌细胞生成素的肌纤维。三色染色和甲苯胺蓝染色支持体内肌纤维形成。乙酰胆碱酯酶的活性提示再生运动单位的功能活性。与未分化的ES细胞移植和注射磷酸盐缓冲盐水（对照损伤）相比，与转染细胞移植的胫前肌的诱发肌电图显示出显着更高的电位。电镜检查证实了体内细胞内肌纤维的形成。结论.将IGFII基因转染ES细胞可用于创伤性肌肉损伤和肌病的移植治疗。
Background. Reconstruction of skeletal muscle tissue is hampered by the lack of availability of functional substitution of the tissue. Methods. Embryonic stem (ES) cells were transfected with the insulin-like growth factor (IGF) II gene and were selected with G418. The resultant cell clones were analyzed regarding their myogenic differentiation in vitro and in vivo. Results. The cells expressed early and late myogenic differentiation markers, including myoD, myogenin, and dystrophin in vitro. They had phosphorylated Akt within the cells, suggesting their activation by the secreted IGFII. Transplantation of the cells to injured anterior tibial muscle of mice significantly improved their motor functions compared to injured mice transplanted with undifferentiated ES cells and injured mice given vehicle alone. The transfected cells adapted to the injured muscle, formed myofibers positive for dystrophin and negative for MyoD and myogenin. Trichrome staining and toluidine blue staining support myofiber formation in vivo. The enzymatic activity of acetylcholine esterase suggested the functional activity of the regenerated motor units. The evoked electromyogram of anterior tibial muscle transplanted with the transfected cells showed significantly higher potentials compared to that transplanted with undifferentiated ES cells and that injected with phosphate-buffered saline (control injury). Electron microscopic examination confirmed the myofiber formation in the cells in vivo. Conclusions. Transfection of IGFII gene into ES cells may be applicable for transplantation therapy of muscle damage due to injury and myopathies.