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  本文關(guān)鍵詞：比較新生大鼠腦部不同部位的神經(jīng)干細(xì)胞被誘導(dǎo)分化為少突膠質(zhì)細(xì)胞的差異　出處：《福建醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 神經(jīng)干細(xì)胞 少突膠質(zhì)細(xì)胞 腦室周圍白質(zhì)軟化 髓鞘

【摘要】： 【目的】 體外培養(yǎng)獲得大腦皮質(zhì)、紋狀體、海馬三個(gè)不同部位來(lái)源的神經(jīng)干細(xì)胞(neural stem cells, NSCs)并對(duì)其進(jìn)行誘導(dǎo)分化,比較三個(gè)不同部位來(lái)源的神經(jīng)干細(xì)胞增殖傳代能力的差別以及定向誘導(dǎo)分化為少突膠質(zhì)細(xì)胞的分化率的差別。為體外培養(yǎng)獲得更多的少突膠質(zhì)細(xì)胞的研究提供實(shí)驗(yàn)依據(jù)。 【方法】 1.分別取新生24h內(nèi)SD大鼠的大腦皮層、海馬、紋狀體部位來(lái)源的神經(jīng)干細(xì)胞在堿性成纖維生長(zhǎng)因子和B27聯(lián)合作用下使其穩(wěn)定增殖,單細(xì)胞克隆實(shí)驗(yàn)檢測(cè)其自我更新能力;免疫熒光染色法對(duì)NSCs標(biāo)志物nestin以及分化為神經(jīng)元和膠質(zhì)細(xì)胞的多向分化能力進(jìn)行鑒定。 2.對(duì)三個(gè)不同部位來(lái)源的神經(jīng)干細(xì)胞進(jìn)行傳代培養(yǎng),CCK8法檢測(cè)不同部位來(lái)源的NSCs的增殖能力,繪制細(xì)胞生長(zhǎng)曲線;比較不同部位來(lái)源的NSCs的增殖及傳代能力差別。 3.分別以胰島素樣生長(zhǎng)因子-1、甲狀腺素、神經(jīng)膠質(zhì)母細(xì)胞瘤培養(yǎng)上清液誘導(dǎo)NSCs分化,并通過激光共聚焦顯微鏡及Western blot法對(duì)成功誘導(dǎo)為OLC的細(xì)胞分別進(jìn)行陽(yáng)性細(xì)胞計(jì)數(shù)與Galc的定量檢測(cè),比較各組神經(jīng)干細(xì)胞定向分化為少突膠質(zhì)細(xì)胞的誘導(dǎo)分化率。 【結(jié)果】 1.采用無(wú)血清培養(yǎng)基培養(yǎng),可獲取高純度的大鼠原代NSCs;nestin標(biāo)志物檢測(cè)陽(yáng)性,自然分化后可分化為少突膠質(zhì)細(xì)胞、神經(jīng)元及星形膠質(zhì)細(xì)胞,免疫熒光染色陽(yáng)性;單細(xì)胞克隆實(shí)驗(yàn)可以獲得單細(xì)胞克隆球。 2.大腦皮質(zhì)來(lái)源的NSCs可以傳代9-12次,海馬來(lái)源的NSCs可以傳代8-11次,紋狀體來(lái)源的NSCs可以傳代6-9次;CCK8法繪制細(xì)胞生長(zhǎng)曲線,統(tǒng)計(jì)分析得:海馬與紋狀體來(lái)源的NSCs的增殖活性無(wú)差異(P㧐0.05);皮質(zhì)與海馬來(lái)源的NSCs增殖活性有差異(P㩳0.05),結(jié)果有統(tǒng)計(jì)學(xué)意義;皮質(zhì)與紋狀體來(lái)源的NSCs增殖活性有差異(P㩳0.05),結(jié)果有統(tǒng)計(jì)學(xué)意義。且隨著傳代次數(shù)的增加,該差異性依然維持。 3.胰島素樣生長(zhǎng)因子-1、甲狀腺素誘導(dǎo)能力無(wú)差異(P㧐0.05);神經(jīng)膠質(zhì)母細(xì)胞瘤培養(yǎng)上清液與胰島素樣生長(zhǎng)因子-1誘導(dǎo)能力有差異(P㩳0.05),結(jié)果有統(tǒng)計(jì)學(xué)意義;神經(jīng)膠質(zhì)母細(xì)胞瘤培養(yǎng)上清液與甲狀腺素誘導(dǎo)能力有差異(P㩳0.05),結(jié)果有統(tǒng)計(jì)學(xué)意義;皮質(zhì)來(lái)源的神經(jīng)干細(xì)胞與海馬及紋狀體來(lái)源的神經(jīng)干細(xì)胞向少突膠質(zhì)細(xì)胞分化率有差異(P㩳0.05),結(jié)果有統(tǒng)計(jì)學(xué)意義; 【結(jié)論】 1.采用無(wú)血清培養(yǎng)基懸浮培養(yǎng),可獲取高純度的具有自我更新及多向分化能力的大鼠原代NSCs。 2.皮質(zhì)來(lái)源的神經(jīng)干細(xì)胞增殖及傳代能力強(qiáng)于海馬、紋狀體部位來(lái)源的神經(jīng)干細(xì)胞。并且體外培養(yǎng)的神經(jīng)干細(xì)胞一直維持該特性。 3.神經(jīng)膠質(zhì)母細(xì)胞瘤培養(yǎng)上清液的誘導(dǎo)神經(jīng)干細(xì)胞向少突膠質(zhì)細(xì)胞分化的能力強(qiáng)于胰島素樣生長(zhǎng)因子-1、甲狀腺素;皮質(zhì)來(lái)源的神經(jīng)干細(xì)胞向少突膠質(zhì)細(xì)胞的分化能力強(qiáng)于海馬、紋狀體部位來(lái)源的神經(jīng)干細(xì)胞。
[Abstract]:[Objective]
Get the cerebral cortex in vitro, striatum, hippocampus three neural stem cells of different origins (neural stem, cells, NSCs) and of its differentiation, three from different parts of neural stem cell proliferation ability difference and differentiation to the differences in the rate of differentiation of oligodendrocytes provide experimental. According to the research get more oligodendrocytes in vitro.
[method]
Hippocampus were obtained from 1. newborn SD rats 24h in the cerebral cortex, striatum derived neural stem cells in basic fibroblast growth factor and B27 under the combined effect of the proliferation of experimental single cell clone detection their self-renewal ability; immunofluorescence staining for NSCs and nestin markers were identified to differentiate into multilineage differentiation the ability of neurons and glial cells.
2., three sources of neural stem cells from different locations were cultured. The proliferation ability of NSCs from different parts was detected by CCK8, and cell growth curve was drawn. The proliferation and passage ability of NSCs from different locations were compared.
3. respectively by insulin-like growth factor -1, thyroxine, NSCs induced differentiation of human glioblastoma supernatant and cultured by confocal microscopy and Western blot method for quantitative OLC induced cells were Galc positive cell counting and detection, comparison of neural stem cells to differentiate into oligodendrocyte differentiation rate cells.
[results]
1. using serum-free culture medium can obtain high-purity primary rat NSCs; nestin markers positive natural differentiation can differentiate into oligodendrocytes, neurons and astrocytes, immunofluorescence staining; single cell clone experiments can get cell clone ball.
2. from the cerebral cortex of NSCs can be passaged 9-12 times, hippocampus derived NSCs can be passaged 8-11 times, from the striatum of NSCs can be passaged 6-9 times; the cell growth curve CCK8 method, statistical analysis: no difference between hippocampus and striatum derived the proliferation activity of NSCs (P? 0.05); NSCs proliferation activity of cortex and hippocampus sources there are differences (P? 0.05), the results were statistically significant; the proliferation of NSCs cortex and striatum derived differences (P? 0.05), the results were statistically significant. With the increase in the number of passages, the differences are still maintained.
3. insulin like growth factor -1, no difference in ability induced by thyroxine (P? 0.05); nerve glioblastoma supernatant and insulin-like growth factor -1 induced ability are different (P? 0.05), the results were statistically significant; and the supernatant induced by thyroxine ability is different in glioblastoma culture (P? 0.05) and the results have statistical significance; cortex derived neural stem cells and hippocampus and striatum derived neural stem cells to oligodendrocyte differentiation rate difference (P? 0.05), the results were statistically significant;
[Conclusion]
1. by suspension culture of serum-free medium, the primary NSCs. of rats with high purity and self renewal and multidirectional differentiation can be obtained.
2., the proliferation and passage ability of cortical derived neural stem cells are stronger than those of hippocampus and striatum. Neural stem cells cultured in vitro maintain this characteristic.
3. human glioblastoma culture to oligodendrocyte differentiation in insulin-like growth factor -1, neural stem cells induced by supernatant of thyroxine; cortical source of neural stem cells to oligodendrocyte differentiation ability in hippocampus, striatum derived neural stem cells.

【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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