体外培养人胚胎生殖干细胞的超微结构及培养体系中细胞因子的表达

发布时间：2018-03-01 10:03

本文关键词： 胚胎生殖干细胞胚胎成纤维细胞细胞因子超微结构　出处：《苏州大学》2005年硕士论文　论文类型：学位论文

【摘要】：目的:检测细胞培养上清液中LIF、SCF和bFGF的水平,探索更完善的人胚胎生殖干细胞(EG细胞)体外培养体系;光镜观察人PGCs,透射电镜观察人EG细胞的超微结构,深入了解人PGCs和人EG细胞的形态学特征。方法:采用组织块培养法体外培养人EG细胞,收集不同生长阶段(第3d和第9d)的细胞培养上清液,用双抗体夹心ABC-ELISA法定量测定其中人LIF、SCF、bFGF的水平;取胚胎两侧生殖腺嵴、中肾嵴常规HE染色,制备切片,在光镜下观察并拍摄照片;收集足量的人EG细胞和胚胎成纤维细胞制备超薄切片,在透射电镜下观察并拍摄照片。结果:第9d培养液中LIF浓度(55.25±6.51pg/ml),高于第3d(46.56±0.89pg/ml),差别有统计意义(P0.05);第9d SCF浓度(90.39±3.98pg/ml),显著高于第3d(78.47±2.03pg/ml),差别有高度统计意义(P0.01);第9d bFGF浓度(26.06±1.73pg/ml),显著高于第3d(22.13±1.08pg/ml),差别有高度统计意义(P0.01)。光镜下可见人PGCs呈索条状排列或零星散布于组织中,细胞大,呈圆形或椭圆形,轮廓清楚。核质比高,细胞核大,着色深,呈圆形或椭圆形,呈偏心位,核仁较明显。透射电镜下观察人EG细胞核质比高,细胞质少,细胞器不发达,
[Abstract]:Objective: to detect the levels of LIF-SCF and bFGF in the supernatant of cell culture, to explore a more perfect culture system of human embryonic reproductive stem cells (EG), to observe human PGCs by light microscope and to observe the ultrastructure of EG cells by transmission electron microscope (TEM). To understand the morphological characteristics of human PGCs and human EG cells. Methods: human EG cells were cultured in vitro by tissue mass culture. The supernatants of different growth stages (3rd and 9th days) were collected. The levels of human LIF-SCFbFGF were measured by double antibody sandwich ABC-ELISA assay, and the gonadal cristae of both sides of embryo were obtained. The mesonephric cristae were stained with HE, sections were prepared, observed and photographed under light microscope, and ultrathin sections were prepared by collecting enough human EG cells and embryonic fibroblasts, and were observed and photographed under transmission electron microscope (TEM). Results: the concentration of LIF in the culture medium on the 9th day was 55.25 卤6.51 PG / ml, which was higher than that in the 3rd day (46.56 卤0.89 PG / ml), the difference was statistically significant (P 0.05), and on the 9th day, the concentration of SCF was 90.39 卤3.98 PG / ml, which was significantly higher than that on the 3rd day (78.47 卤2.03 PG / ml), and the bFGF concentration on the 9th day was 26.06 卤1.73 PG / ml, which was significantly higher than that on the 3rd day (22.13 卤1.08pg / ml). Under the microscope, we can see that the human PGCs is arranged in the form of cable or scattered in the tissue. The cells were large, round or oval, with clear outline, high ratio of nucleus to cytoplasm, large nucleus, deep staining, round or ellipse, eccentric position, obvious nucleolus, high cytoplasm ratio and less cytoplasm of human EG observed by transmission electron microscope. The organelles are underdeveloped,
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R321

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