论文部分内容阅读
目的:观察并探讨携人胰岛素样生长因子-1(hIGF-1)基因的成肌细胞移植入雄性C3H小鼠体内后的存活、转归以及移植后hIGF-1基因的表达。方法:84只雄性C3H小鼠随机(20~30g,7~11w)分为A组(正常小鼠转基因细胞移植组)、B组(正常小鼠空白成肌细胞移植组)、C组(受伤小鼠转基因细胞移植组)和D组(受伤小鼠空白成肌细胞移植组),每组20只,另4只作正常对照。A、B两组分别于右侧腓肠肌中段注射转基因成肌细胞或空白成肌细胞;C、D两组以重力打击造成小鼠右侧腓肠肌中段钝挫伤,伤后第3天分别于致伤局部注射转基因成肌细胞或空白成肌细胞。注射细胞后第2、5、10、20、30天,各组随机抽取4只小鼠处死,取右侧腓肠肌中段检测,Br-dU免疫组化染色检测外源细胞在体内的存活情况;4组另行hIGF-1免疫组化染色及实时PCR检测外源转基因细胞在体内表达hIGF-1情况。结果:各组小鼠均有BrdU免疫组化阳性染色,A、C两组均有hIGF-1mRNA表达及hIGF-1分泌,B、D两组未检测到hIGF-1mRNA表达及hIGF-1分泌。结论:携hIGF-1基因的成肌细胞移植入正常及钝挫伤小鼠体内后,可存活一段时间,并能稳定地分泌hIGF-1因子。
OBJECTIVE: To investigate the survival, prognosis and the expression of hIGF-1 gene after transplantation of myoblasts carrying human insulin like growth factor-1 (hIGF-1) gene into male C3H mice. Methods: Eighty-four male C3H mice were randomly divided into group A (normal mouse transgenic cell transplantation group), group B (normal mouse blank myoblast transplantation group), group C (injured Mice transgenic mice transplantation group) and D group (injured mice blank myoblast transplantation group), each group of 20, the other four as a normal control. Groups A and B were injected with either transgenic myoblasts or blank myoblasts respectively in the right gastrocnemius muscle. C and D groups caused blunt contusions in the middle part of the right gastrocnemius muscle by gravity. On the 3rd day after injury, Transgenic myoblasts or blank myoblasts were injected. At 2, 5, 10, 20 and 30 days after injection, 4 mice were randomly selected in each group and sacrificed. The right middle gastrocnemius muscle was detected and Br-dU immunohistochemical staining was used to detect the survival of exogenous cells in vivo. Group hIGF-1 immunohistochemical staining and real-time PCR detection of foreign genetically modified cells in vivo expression hIGF-1 situation. Results: Immunohistochemical staining of BrdU in each group showed positive staining. HIGF-1mRNA and hIGF-1 secretion were found in groups A and C, while hIGF-1mRNA and hIGF-1 secretion were not detected in groups B and D. CONCLUSION: The hIGF-1 gene myoblasts transplanted into normal and blunt trauma mice can survive for some time and can stably secrete hIGF-1 factor.