小鼠四倍体胚胎凋亡检测及嵌合胚胎制作

Apoptosis detection of tetraploid embryos and production on chimeric embryos

四倍体胚胎补偿技术,特别是ES细胞和iPS细胞功能验证是已成为目前干细胞研究领域热点。本试验旨在通过提高四倍体胚胎与ES细胞形成嵌合胚胎效率,为构建转基因动物模型搭建技术平台。本研究利用电融合法制作小鼠四倍体胚胎,并采用TUNEL法检测四倍体胚胎凋亡情况以对其进行质量评估,同时通过四倍体胚胎补偿法和聚合法,将小鼠四倍体胚胎与带有GFP标记ES细胞制作成嵌合胚胎,从而能够为快速、简便获得完全ES细胞发育而来生物个体提供重要技术支持,并为基因功能研究和疾病动物模型构建提供一定研究基础。

Tetraploid embryo complementation, especially ES and iPS have been a research hot spot now. The purpose of this study is to build a platform for the construction of transgenic animal models. Electric fusion method was used to prepare the mouse tetraploid embryos, TUNEL assay was used for the detection of cell apoptosis. The compensation method and polymerization were used to produce tetraploid embryos with GFP labeled and ES cells into chimeric embryos of the mouse. This method can be easy and fast access to individuals completely developed from ES cells, and provided an effective way to study gene function and construction of transgenic animal models.