【摘要】： 第一部分骨髓間充質(zhì)干細胞的多向分化潛能 第一章體外誘導(dǎo)成年大鼠骨髓間充質(zhì)干細胞分化為5羥色胺敏感性神經(jīng)元 目的:體外誘導(dǎo)成年大鼠骨髓間充質(zhì)干細胞分化為具有神經(jīng)元表型和部分功能的細胞。 方法:將取自6-8周成年SD大鼠股、脛骨的細胞貼壁傳代培養(yǎng)至6-10代,采用有限稀釋法對細胞作單克隆純化。在全反式視黃酸預(yù)誘導(dǎo)24小時后,HBSS洗去含血清培養(yǎng)液,換用改良的Woodbury長時程神經(jīng)元誘導(dǎo)液培養(yǎng)細胞,3、6、9、12、24、48、72、96小時后顯微鏡下觀察細胞形態(tài)變化;細胞免疫染色檢測神經(jīng)元標志物神經(jīng)元特異性烯醇化酶、神經(jīng)絲M+H、5-羥色胺的表達;并用胞內(nèi)鈣代謝測定法測定誘導(dǎo)前后細胞對5-HT的功能反應(yīng)。 結(jié)果:改良神經(jīng)元誘導(dǎo)液處理3小時后,細胞開始表現(xiàn)神經(jīng)元的形態(tài)特征,細胞折光性增強,形成收縮的雙極或多極胞體和細長突起。細胞可以維持神經(jīng)元樣存活72小時以上。誘導(dǎo)5小時后,對免疫染色
[Abstract]:The first part is the multidirectional differentiation potential of bone marrow mesenchymal stem cells. Chapter 1 inducing adult rat bone marrow mesenchymal stem cells to differentiate into serotonin sensitive neurons in vitro Aim: to induce adult rat bone marrow mesenchymal stem cells to differentiate into cells with neuronal phenotype and partial function in vitro. Methods: the cells taken from the strands of adult SD rats for 6 weeks and 8 weeks were cultured to 6 脳 10 generations, and the cells were purified by finite dilution method. After 24 hours of all-trans Retinic acid pretreatment, HBSS washed away the serum culture medium and replaced it with the modified Woodbury long-term neuron induction solution. The cells were cultured in 3, 6, 9, 12, 24, 48, 72, 96 hours, and the morphological changes of the cells were observed under microscope. The expression of neuron-specific enolase and 5-hydroxytryptamine in nerve filament was detected by cellular immunostaining, and the functional response of cells to 5-HT before and after induction was measured by intracellular calcium metabolism assay. Results: after 3 hours of modified neuron induction solution treatment, the cells began to show the morphological characteristics of neurons, the refractive index of the cells was enhanced, and the contractile bipolar or multipolar cell bodies and slender processes were formed. Cells can maintain neuron-like survival for more than 72 hours. After 5 hours of induction, immunostaining
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R329

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