本文關(guān)鍵詞： 臍帶 間充質(zhì)干細(xì)胞 神經(jīng)細(xì)胞 分化 誘導(dǎo) 丹參　出處：《汕頭大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：研究人臍帶間充質(zhì)干細(xì)胞(MSCs)的體外分離、純化、擴(kuò)增及丹參誘導(dǎo)MSCs向神經(jīng)元樣細(xì)胞分化，，為神經(jīng)細(xì)胞移植、臍帶組織庫的建立及中藥丹參作為神經(jīng)誘導(dǎo)劑提供理論依據(jù)。方法：無菌條件下收集剖宮產(chǎn)新生兒臍帶，組織塊培養(yǎng)法獲取MSCs，進(jìn)行培養(yǎng)。丹參誘導(dǎo)人臍帶來源的MSCs向神經(jīng)細(xì)胞分化；用免疫細(xì)胞化學(xué)方法檢測(cè)神經(jīng)細(xì)胞相關(guān)蛋白標(biāo)記的表達(dá)；用光鏡、掃描電子顯微鏡監(jiān)測(cè)丹參誘導(dǎo)后細(xì)胞形態(tài)改變；透射電子顯微鏡觀察丹參誘導(dǎo)后細(xì)胞超微結(jié)構(gòu)改變；用共聚焦顯微鏡分析丹參誘導(dǎo)前后細(xì)胞內(nèi)DNA、RNA含量改變及神經(jīng)干細(xì)胞的標(biāo)記巢蛋白(nestin)，神經(jīng)元的蛋白標(biāo)記β-Ⅲ類神經(jīng)微管(β-TubulinⅢ)表達(dá)含量變化。用RT-PCR檢測(cè)細(xì)胞神經(jīng)相關(guān)基因pleiotrophin和神經(jīng)干細(xì)胞的標(biāo)記巢蛋白(nestin)的表達(dá)。RT-PCR檢測(cè)證實(shí)，經(jīng)丹參誘導(dǎo)后的MSCs表達(dá)神經(jīng)干細(xì)胞相關(guān)基因Nestin，誘導(dǎo)前和誘導(dǎo)后的MSCs均表達(dá)神經(jīng)細(xì)胞基因Pleiotrophin，誘導(dǎo)后的表達(dá)明顯增強(qiáng)。結(jié)果：從人臍帶華爾通膠分離、培養(yǎng)的貼壁細(xì)胞，體外生長(zhǎng)形態(tài)類似于成纖維細(xì)胞，可以維持在未分化狀態(tài)穩(wěn)定增殖，體外增殖超過10代，細(xì)胞凍存一個(gè)月后
[Abstract]:Objective: to study the isolation, purification, amplification and differentiation of MSCs into neuron-like cells from human umbilical cord mesenchymal stem cells (MSCs) in vitro. The establishment of umbilical cord tissue bank and traditional Chinese medicine Salvia miltiorrhiza as a nerve inducer to provide theoretical basis. Methods: the umbilical cord of newborn born by cesarean section was collected under aseptic condition and MSCs was obtained by tissue mass culture. Salvia miltiorrhiza induced the differentiation of MSCs from human umbilical cord into neural cells. The expression of neuron-associated protein was detected by immunocytochemistry. The morphological changes of cells induced by Salvia miltiorrhiza were monitored by light microscope and scanning electron microscope. The ultrastructure of cells induced by Salvia miltiorrhiza was observed by transmission electron microscope. The changes of DNA RNA content and the labeled nestin of neural stem cells were analyzed by confocal microscope before and after salvia miltiorrhiza induction. Protein labeling of Neuron 尾-鈪

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