标准化嵌合体制备体系的研究

目的:完善和规范基因剔除小鼠技术体系的关键技术环节,建立一套高效的嵌合体制备体系。方法:优化胚胎干细胞(ES细胞)的基本培养条件;应用条件培养液筛选、富集高嵌合潜能的ES细胞;成熟嵌合体的制备技术;改变胚胎的移植方式,改善受体的生理状态。结果:ES细胞基本培养条件的优化及条件培养液的筛选保持了ES细胞的整体高嵌合潜能,嵌合体制备技术得以成熟,胚胎移植方式的改变提高了移植胚胎的产仔率,这些措施大大提高了嵌合体的制备效率。结论:通过对基因剔除小鼠技术体系的关键技术环节进行优化和改进,建立了一套高效的嵌合体制备程序,为基因剔除小鼠服务体系的开展打下了坚实的基础。

Study on Standardization System for Chimeras Production

Objective:To establish a highly efficient production system of chimeras by perfecting and normalizing the key steps of technology system of gene knockout mice.Methods:To optimize the basic cultivation conditions of ES cells;to screen and enrich the high chimeric potential of ES cells by using conditional cultivation medium;to improve the production efficiency of chimeras by maturing correlated technique;to improve the state of recipient mice by changing the method of embryo transplantation.Results:High chimeric potency of ES cells was maintained by optimization of basic cultivation conditions and use of conditional cultivation medium.The production technology of chimeras was mature,and the fetiferous rate was improved by changing the method of embryo transplantation.These measures increased the production efficiency of chimera remarkably.Conclusion:This study established a highly efficient production system of chimeras by perfecting and normalizing the key steps of technology system of gene knockout mice,which has laid a solid foundation for offering the serve of gene knockout mice technology.