摘要
目的建立一种有效的研究成纤维细胞体内转归的新模型。方法以绿色荧光蛋白转基因小鼠成纤维细胞为研究对象，将其以每毫升20×10。个细胞浓度注射1个点，每点0．15ml注射到裸鼠头皮下，注射后形成直径1cm的圆丘。1周后通过活体荧光检测仪，冰冻切片荧光显微镜及GFP蛋白免疫组化检测，观察其在裸鼠头部皮下存在的数量和分布情况。结果GFP小鼠成纤维细胞裸鼠头皮下注射1周后，活体荧光定性分析可见荧光以注射点为中心成同心圆分布，并且荧光区域大于注射区域，荧光强度不均匀；活体荧光信号定量分析可见其平均荧光灰度强度值：92677419692（Ph／mm^2·s）；冰冻切片荧光显微镜可发现注射部位的细胞发出明显的绿色荧光；GFP蛋白免疫组化染色可发现注射部位明显的GFP蛋白存在。结论通过绿色荧光蛋白转基因小鼠成纤维细胞体内注射，活体荧光检测仪，冰冻切片荧光显微镜及GFP蛋白免疫组化观察其在裸鼠头部皮下存在的数量和分布情况的方法，建立一种有效的研究成纤维细胞体内转归的模型。
Objective To establish a new model of reasearch on prognosis of fibroblast injected in-vivo. Methods Fibroblasts were derived from Green fluorescent protein （GFP） transgenic mice. After 3 passages of culture, cells were harvested and injected subcutaneously into the scalp of nude mice at density of 20×10^6cells/ml. At 1 week, the head of the nude mice were examined by NightOWL LB983 in vivo imaging system to characterize the distribution and the survival rate of GFP-labeled fibroblasts. The fluorescence intensity of the fibroblasts was analyzed statistically. The head skin of nude mice within lcm around the injection size was dis- sected and characterizedby frozen section. The distribution and survival of GFP-labeled fibroblasts were further examined under the fluorescence microscope. The tissue sections were also analyzed by histological examination and GFP immunohistochemistry to confirm the prognosis of fibroblast cell at the injection site. Results The re- sult indicated that we discovered obviously the fibroblast at 1 week. Conclusion We fund a new study model to investigate the prognosis of fibroblast after injection in vivo.
出处
《中国美容整形外科杂志》
CAS
2012年第2期122-125,共4页
Chinese Journal of Aesthetic and Plastic Surgery
基金
北京市科委科技计划重大项目（12090800046609003）
关键词
GFP小鼠
成纤维细胞
皮下注射
转归
GFP transgenic mice
Fibroblast
Hypodermic injection
Prognosis
