人胚胎生殖干细胞的生物学鉴定

发布时间：2018-04-22 09:19

本文选题：阶段特异性胚胎抗原 + 胚胎生殖干细胞　；参考：《苏州大学》2005年硕士论文

【摘要】：目的:组织块培养法培养人胚胎生殖干细胞(EG 细胞),通过检测细胞所表达的阶段特异性胚胎抗原1 和3(stage-specific embryonic antigen-1、3 ,SSEA-1、SSEA-3),对未分化的人EG 细胞进行特异性的生物学鉴定。方法:取5—9 周流产人胚胎的背侧肠系膜和生殖腺嵴,进行组织块体外培养,在培养液中不添加细胞因子,以同源胚胎的成纤维细胞作为饲养层细胞,利用免疫细胞化学的方法,对组织块培养的细胞表达的SSEA-1、SSEA-3 进行检测;取5-9 周人胚胎生殖腺嵴做石蜡切片,采用免疫组织化学的方法,对生殖腺嵴中的原始生殖细胞(primordial germ cells,PGCs)进行SSEA-1、SSEA-3 的检测。结果:石蜡切片免疫组织化学方法显示,5-9 人周胚胎生殖腺嵴中有大量成条索状排列、SSEA-1、SSEA-3 呈阳性表达的PGCs;体外培养过程中,在胚胎成纤维细胞上生长的单个EG 样细胞及其集落,均表达SSEA-1、SSEA-3。结论:本实验组织块培养所获细胞来源于PGCs,为未分化的人EG 细胞。
[Abstract]:Objective: to culture human embryonic reproductive stem cells (EG) cells by tissue culture, and to identify the undifferentiated human EG cells by detecting the expression of phase specific embryonic antigen 1 and 3(stage-specific embryonic antigen-1p3 SSEA-1SSEA-3. Methods: the dorsal mesenteric and gonadal crest of human embryo aborted for 5-9 weeks were cultured in vitro with tissue mass. The fibroblasts of homologous embryos were used as feeder layer cells without cytokines in the culture medium. The expression of SSEA-1and SSEA-3 was detected by immunocytochemistry, and the gonadal cristae of human embryos of 5-9 weeks were taken as paraffin sections, and the expression of SSEA-3 was detected by immunohistochemical method. The primordial germ cells of primordial germ cells were detected by SSEA-1and SSEA-3 in gonadal crest. Results: the paraffin section immunohistochemical method showed that there were a large number of PGCspositive expression of SSEA-1and SSEA-3 in the gonadal crest of 5-9 human periembryonic embryos, and a single EG-like cell and its colony were grown on the embryonic fibroblasts during the process of culture in vitro. Both expressed SSEA-1 and SSEA-3. Conclusion: the cells derived from PGCswere undifferentiated human EG cells.
【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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