【摘要】：目的探討小鼠孤雌胚胎干細(xì)胞在體外不經(jīng)過擬胚體(EB)階段,直接誘導(dǎo)分化為神經(jīng)細(xì)胞的可行性。方法采用階段誘導(dǎo)的方法,首先由孤雌胚胎干細(xì)胞向神經(jīng)前體細(xì)胞定向分化,通過巢蛋白(nestin)免疫熒光細(xì)胞化學(xué)染色對神經(jīng)前體細(xì)胞進(jìn)行鑒定。在此基礎(chǔ)上,撤除絲裂原,加入5%胎牛血清,觀察已分化的神經(jīng)前體細(xì)胞能否進(jìn)一步分化為神經(jīng)細(xì)胞,并對分化出的細(xì)胞進(jìn)行免疫熒光細(xì)胞化學(xué)鑒定。結(jié)果經(jīng)選擇培養(yǎng)基培養(yǎng)3 d后的孤雌干細(xì)胞絕大多數(shù)可誘導(dǎo)為神經(jīng)前體細(xì)胞,nestin呈陽性。加入血清進(jìn)一步誘導(dǎo),可分化出β-Ⅲ-tubline陽性的神經(jīng)細(xì)胞。結(jié)論小鼠孤雌胚胎干細(xì)胞在體外不經(jīng)EB培養(yǎng)階段,可直接誘導(dǎo)分化為神經(jīng)細(xì)胞,并且前期可得到大量的神經(jīng)前體細(xì)胞。
[Abstract]:Objective to investigate the feasibility of differentiation of mouse parthenogenetic embryonic stem cells into neural cells without embryoid (EB) in vitro. Methods the neural progenitor cells were differentiated from parthenogenetic embryonic stem cells to neural precursor cells by the method of stage induction. The neural progenitor cells were identified by (nestin) immunofluorescence cytochemical staining. On this basis, the mitogen was removed and 5% fetal bovine serum was added to observe whether the differentiated neural precursor cells could be further differentiated into nerve cells, and the differentiated cells were identified by immunofluorescence cytochemistry. Results the majority of parthenogenetic stem cells cultured in selected medium for 3 days could be induced to be neural precursor cells and nestin was positive. The 尾-鈪，

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