本文關(guān)鍵詞： 臍帶 間充質(zhì)干細胞 神經(jīng)細胞 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)干細胞(MSCs)的體外分離、純化、擴增及丹參誘導(dǎo)MSCs向神經(jīng)元樣細胞分化，，為神經(jīng)細胞移植、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，進行培養(yǎng)。丹參誘導(dǎo)人臍帶來源的MSCs向神經(jīng)細胞分化；用免疫細胞化學(xué)方法檢測神經(jīng)細胞相關(guān)蛋白標記的表達；用光鏡、掃描電子顯微鏡監(jiān)測丹參誘導(dǎo)后細胞形態(tài)改變；透射電子顯微鏡觀察丹參誘導(dǎo)后細胞超微結(jié)構(gòu)改變；用共聚焦顯微鏡分析丹參誘導(dǎo)前后細胞內(nèi)DNA、RNA含量改變及神經(jīng)干細胞的標記巢蛋白(nestin)，神經(jīng)元的蛋白標記β-Ⅲ類神經(jīng)微管(β-TubulinⅢ)表達含量變化。用RT-PCR檢測細胞神經(jīng)相關(guān)基因pleiotrophin和神經(jīng)干細胞的標記巢蛋白(nestin)的表達。RT-PCR檢測證實，經(jīng)丹參誘導(dǎo)后的MSCs表達神經(jīng)干細胞相關(guān)基因Nestin，誘導(dǎo)前和誘導(dǎo)后的MSCs均表達神經(jīng)細胞基因Pleiotrophin，誘導(dǎo)后的表達明顯增強。結(jié)果：從人臍帶華爾通膠分離、培養(yǎng)的貼壁細胞，體外生長形態(tài)類似于成纖維細胞，可以維持在未分化狀態(tài)穩(wěn)定增殖，體外增殖超過10代，細胞凍存一個月后
[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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