【摘要】：背景與目的 胚胎干細(xì)胞(embryonic stem cells，ESCs)是從植入前囊胚的內(nèi)細(xì)胞團(tuán)(inner cell mass，ICM)或原始生殖細(xì)胞(primordial germ cells，PGCs)分離和克隆的具有全能性細(xì)胞。理論上，胚胎干細(xì)胞可保持未分化的狀態(tài)在體外無(wú)限傳代培養(yǎng)。在適當(dāng)?shù)沫h(huán)境中又可以分化成包括體細(xì)胞、生殖細(xì)胞在內(nèi)的任何類(lèi)型的細(xì)胞，，并具有正常的核型。可以利用核移植技術(shù)建立核移植胚胎干細(xì)胞(nuclear transfer embryonic stem cells．ntES)系，利用胚胎干細(xì)胞制備嵌合體動(dòng)物，對(duì)胚胎干細(xì)胞進(jìn)行基因改造等研究。胚胎干細(xì)胞的發(fā)現(xiàn)為研究細(xì)胞分化，物種發(fā)育機(jī)制，闡明基因功能，篩選藥物等開(kāi)辟了廣闊的空間。人胚胎干細(xì)胞的建系為人類(lèi)醫(yī)學(xué)帶來(lái)了革命性的改變。人胚胎干細(xì)胞相關(guān)的各種定向分化研究、組織工程的進(jìn)展為世界各個(gè)研究領(lǐng)域的專(zhuān)家所矚目。但是，胚胎干細(xì)胞對(duì)培養(yǎng)條件要求苛刻，容易自發(fā)分化，難以維持正常的二倍體核型。眾多在美國(guó)國(guó)立衛(wèi)生研究院(national institutes of health，NIH)注冊(cè)的人胚胎干細(xì)胞系不能長(zhǎng)期保持發(fā)售。因此，胚胎干細(xì)胞的穩(wěn)定建系和未分化機(jī)制研究成為熱點(diǎn)和難點(diǎn)。本研究嘗試從昆明白小鼠囊胚中分離ES細(xì)胞，探討不同的受孕方式、不同培養(yǎng)液組成、不同囊胚內(nèi)細(xì)胞團(tuán)的分離方式、不同的ICM評(píng)分對(duì)昆明小鼠胚胎干細(xì)胞分離的影響，并對(duì)分離獲得的鼠胚胎干細(xì)胞的部分特性作了初步鑒定。為胚胎干細(xì)胞相關(guān)的實(shí)驗(yàn)鋪墊了道路，為進(jìn)一步建立人ES細(xì)胞系和后續(xù)研究奠定基礎(chǔ)。 材料與方法 1．囊胚均來(lái)自于孕3.5天的雌性昆明小白鼠。將囊胚和鼠胚胎成纖維細(xì)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
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R321

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