小鼠胚胎干细胞系的建立及鉴定的初步探讨
发布时间：2018-08-18 12:50
【摘要】：背景与目的 胚胎干细胞(embryonic stem cells，ESCs)是从植入前囊胚的内细胞团(inner cell mass，ICM)或原始生殖细胞(primordial germ cells，PGCs)分离和克隆的具有全能性细胞。理论上，胚胎干细胞可保持未分化的状态在体外无限传代培养。在适当的环境中又可以分化成包括体细胞、生殖细胞在内的任何类型的细胞，，并具有正常的核型。可以利用核移植技术建立核移植胚胎干细胞(nuclear transfer embryonic stem cells．ntES)系，利用胚胎干细胞制备嵌合体动物，对胚胎干细胞进行基因改造等研究。胚胎干细胞的发现为研究细胞分化，物种发育机制，阐明基因功能，筛选药物等开辟了广阔的空间。人胚胎干细胞的建系为人类医学带来了革命性的改变。人胚胎干细胞相关的各种定向分化研究、组织工程的进展为世界各个研究领域的专家所瞩目。但是，胚胎干细胞对培养条件要求苛刻，容易自发分化，难以维持正常的二倍体核型。众多在美国国立卫生研究院(national institutes of health，NIH)注册的人胚胎干细胞系不能长期保持发售。因此，胚胎干细胞的稳定建系和未分化机制研究成为热点和难点。本研究尝试从昆明白小鼠囊胚中分离ES细胞，探讨不同的受孕方式、不同培养液组成、不同囊胚内细胞团的分离方式、不同的ICM评分对昆明小鼠胚胎干细胞分离的影响，并对分离获得的鼠胚胎干细胞的部分特性作了初步鉴定。为胚胎干细胞相关的实验铺垫了道路，为进一步建立人ES细胞系和后续研究奠定基础。 材料与方法 1．囊胚均来自于孕3.5天的雌性昆明小白鼠。将囊胚和鼠胚胎成纤维细胞
[Abstract]:Background & objective embryonic stem cells (embryonic stem cells) were isolated and cloned from the (inner cell mass of the preimplantation blastocyst or the primordial germ cell (primordial germ cells. Having totipotent cells. In theory, embryonic stem cells can remain undifferentiated and cultured indefinitely in vitro. In the appropriate environment, they can be differentiated into any type of cells, including somatic cells and germ cells, and have normal karyotype. Nuclear transfer technology can be used to establish nuclear transfer embryonic stem cells (nuclear transfer embryonic stem cells.ntES) lines, embryonic stem cells to prepare chimeric animals, embryonic stem cells for genetic modification, and so on. The discovery of embryonic stem cells opens up a wide space for the study of cell differentiation, the mechanism of species development, the elucidation of gene function and the screening of drugs. The establishment of human embryonic stem cells has revolutionized human medicine. The progress of tissue engineering in the research of human embryonic stem cells related to directional differentiation has attracted much attention from experts all over the world. However, embryonic stem cells are difficult to maintain normal diploid karyotype. A large number of human embryonic stem cell lines registered with the National Institutes of Health (national institutes of) cannot remain on sale for long periods of time. Therefore, the stability and undifferentiation mechanism of embryonic stem cells have become a hot and difficult point. In this study, es cells were isolated from the blastocysts of Kunming white mice, and the effects of different conception modes, different media composition, different cell mass isolation methods and different ICM scores on the isolation of mouse embryonic stem cells were investigated. Some characteristics of mouse embryonic stem cells were preliminarily identified. It paved the way for embryonic stem cell-related experiments and laid the foundation for further establishment of human es cell line and further research. Materials and methods 1. Blastocysts were obtained from female Kunming mice on day 3.5. Fibroblasts from blastocysts and mouse embryos
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R321
[Abstract]:Background & objective embryonic stem cells (embryonic stem cells) were isolated and cloned from the (inner cell mass of the preimplantation blastocyst or the primordial germ cell (primordial germ cells. Having totipotent cells. In theory, embryonic stem cells can remain undifferentiated and cultured indefinitely in vitro. In the appropriate environment, they can be differentiated into any type of cells, including somatic cells and germ cells, and have normal karyotype. Nuclear transfer technology can be used to establish nuclear transfer embryonic stem cells (nuclear transfer embryonic stem cells.ntES) lines, embryonic stem cells to prepare chimeric animals, embryonic stem cells for genetic modification, and so on. The discovery of embryonic stem cells opens up a wide space for the study of cell differentiation, the mechanism of species development, the elucidation of gene function and the screening of drugs. The establishment of human embryonic stem cells has revolutionized human medicine. The progress of tissue engineering in the research of human embryonic stem cells related to directional differentiation has attracted much attention from experts all over the world. However, embryonic stem cells are difficult to maintain normal diploid karyotype. A large number of human embryonic stem cell lines registered with the National Institutes of Health (national institutes of) cannot remain on sale for long periods of time. Therefore, the stability and undifferentiation mechanism of embryonic stem cells have become a hot and difficult point. In this study, es cells were isolated from the blastocysts of Kunming white mice, and the effects of different conception modes, different media composition, different cell mass isolation methods and different ICM scores on the isolation of mouse embryonic stem cells were investigated. Some characteristics of mouse embryonic stem cells were preliminarily identified. It paved the way for embryonic stem cell-related experiments and laid the foundation for further establishment of human es cell line and further research. Materials and methods 1. Blastocysts were obtained from female Kunming mice on day 3.5. Fibroblasts from blastocysts and mouse embryos
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R321
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