本文選題：神經(jīng)干細(xì)胞 + 膠質(zhì)瘤　； 參考：《蘇州大學(xué)》2005年碩士論文

【摘要】：【目的】探索人胚神經(jīng)干細(xì)胞及膠質(zhì)瘤腫瘤干細(xì)胞體外分離培養(yǎng)的條件，并觀察比較其在體外分離培養(yǎng)的生長(zhǎng)特性。用CD133免疫磁珠分選陽(yáng)性細(xì)胞后為進(jìn)一步研究提供材料。 【方法】神經(jīng)干細(xì)胞：取米非司酮流產(chǎn)8-12周胎齡的人胚胎，從胎腦皮層分離組織，，采用胰蛋白酶消化和機(jī)械吹打的方法獲得單細(xì)胞懸液。采用無(wú)血清培養(yǎng)基(DMEM／F12)，加入EGF和bFGF(20ng／ml)細(xì)胞因子和N_2添加劑刺激細(xì)胞增殖，從而進(jìn)行體外擴(kuò)增培養(yǎng)、連續(xù)傳代。添加10％胎牛血清誘導(dǎo)分化，免疫細(xì)胞化學(xué)鑒定神經(jīng)干細(xì)胞(Nestin)和分化成熟的神經(jīng)元(NSE)、星形膠質(zhì)細(xì)胞(GFAP)及少突膠質(zhì)細(xì)胞(CNP)。膠質(zhì)瘤腫瘤干細(xì)胞：將SHG44細(xì)胞系和人腦膠質(zhì)瘤手術(shù)標(biāo)本采用胰蛋白酶消化獲得單細(xì)胞，用CD133免疫磁珠分離陽(yáng)性細(xì)胞、之后分別應(yīng)用無(wú)血清培養(yǎng)基(DMEM／F12，添加bFGF、EGF)培養(yǎng)。形成腫瘤球后用添加10％胎牛血清誘導(dǎo)分化，免疫細(xì)胞化學(xué)染色檢測(cè)腫瘤球細(xì)胞及其分化細(xì)胞中的Nestin、NSE、GFAP陽(yáng)性細(xì)胞。神經(jīng)干細(xì)胞和腫瘤干細(xì)胞在下一步應(yīng)用之前均行CD133免疫磁珠復(fù)選。同一時(shí)間點(diǎn)行流式細(xì)胞儀檢測(cè)細(xì)胞周期，比較增殖活力。 【結(jié)果】從人胎腦分離的細(xì)胞培養(yǎng)在含有EGF和bFGF的無(wú)血清培
[Abstract]:[objective] to explore the conditions of isolation and culture of human embryonic neural stem cells and glioma tumor stem cells in vitro, and to observe and compare the growth characteristics of human embryonic neural stem cells and glioma tumor stem cells in vitro.CD133 immunomagnetic beads were used to separate positive cells for further study.[methods] Neural stem cells (NSCs): single cell suspension was obtained by trypsin digestion and mechanical blow from human embryos of 8-12 weeks of gestational age induced by mifepristone.EGF and bFGF20 ng / ml) cytokines and N2Additives were added to the serum-free medium DMEM / F12 to stimulate the proliferation of the cells, and then the cells were amplified and cultured in vitro and subcultured continuously.Addition of 10% fetal bovine serum to induce differentiation, immunocytochemical identification of neural stem cells (Nestin) and the differentiation of mature neurons, astrocytes (GFAP) and oligodendrocytes (CNP).Glioma tumor stem cells: SHG44 cell lines and human glioma specimens were digested with trypsin to obtain single cells, positive cells were isolated by CD133 immunomagnetic beads, and then cultured in serum-free medium (DMEM / F12) supplemented with bFGFG (EGF).After the tumor ball was formed, 10% fetal bovine serum was added to induce the differentiation. The positive cells of the tumor ball cells and their differentiated cells were detected by immunocytochemical staining.Neural stem cells (NSCs) and tumor stem cells (NSCs) were tested with CD133 immunomagnetic beads before further application.The cell cycle was measured by flow cytometry at the same time point, and the proliferative activity was compared.[results] cells isolated from human fetal brain were cultured in serum-free culture containing EGF and bFGF.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329.2;R739.4

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