本文選題：骨髓基質(zhì)干細(xì)胞 + 褪黑素　； 參考：《中國醫(yī)科大學(xué)》2005年博士論文

【摘要】：目的 神經(jīng)干細(xì)胞的發(fā)現(xiàn)及其分離鑒定、培養(yǎng)技術(shù)的建立,使神經(jīng)細(xì)胞不可再生的傳統(tǒng)理論面臨巨大挑戰(zhàn),并為神經(jīng)系統(tǒng)疾病的治療提供了新的思路。但神經(jīng)干細(xì)胞數(shù)量少、分布分散,體外培養(yǎng)時取材困難,無免疫原性神經(jīng)干細(xì)胞系建立困難,以及異體移植的種屬差異和社會學(xué)、倫理學(xué)等方面的問題都極大地限制了神經(jīng)干細(xì)胞的應(yīng)用。成體動物的骨髓中存在一定數(shù)量的骨髓基質(zhì)干細(xì)胞(MSCs),骨髓基質(zhì)干細(xì)胞在適當(dāng)條件下可以向神經(jīng)干細(xì)胞、神經(jīng)元、和神經(jīng)膠質(zhì)細(xì)胞方向分化。MSCs誘導(dǎo)分化為神經(jīng)元樣細(xì)胞的研究工作已有一定突破,但誘導(dǎo)形成的神經(jīng)元樣細(xì)胞存在著存活時間不長,24小時后大多死亡的問題。故如何延長MSCs源性的神經(jīng)元樣細(xì)胞的壽命是極為重要的。另一方面MSCs移植治療缺血性腦損傷雖己取得一定的進(jìn)展,但MSCs移植后在體內(nèi)環(huán)境下分化為神經(jīng)膠質(zhì)細(xì)胞的比率較大而分化為神經(jīng)細(xì)胞的比率較小,所以如將骨髓基質(zhì)干細(xì)胞在體外分化為神經(jīng)元樣細(xì)胞,再行細(xì)胞移植將會有事半功倍的效果。 本實(shí)驗(yàn)應(yīng)用體外細(xì)胞培養(yǎng)技術(shù)將大鼠骨髓基質(zhì)干細(xì)胞分離培養(yǎng)和純化,測定MSCs的細(xì)胞活力、生長曲線和貼壁率,用形態(tài)學(xué)方法鑒定培養(yǎng)的骨髓基質(zhì)干細(xì)胞。將培養(yǎng)成功的骨髓基質(zhì)干細(xì)胞在體外傳代、擴(kuò)增,以得到一定數(shù)量的細(xì)胞。以褪黑素(MT)和堿性成纖維細(xì)胞生長因子(bFGF)予以干預(yù),以期延長誘導(dǎo)的神經(jīng)元樣細(xì)胞的存活時間,并應(yīng)用生存時間較長的MSCs源性神經(jīng)元樣細(xì)胞,對局灶性腦缺血大鼠模型(MCAO)進(jìn)行細(xì)胞移植。觀察植入細(xì)胞的分布及存活情況,并探討細(xì)胞植入后對MCAO大鼠腦神經(jīng)元C-myc、Erk蛋白及其mRNA表達(dá)的影響。
[Abstract]:Objective the discovery, isolation and identification of neural stem cells and the establishment of culture techniques have challenged the traditional theory of nerve cell non-regeneration. It also provides a new idea for the treatment of nervous system diseases. However, the number of neural stem cells is small, the distribution of neural stem cells is scattered, it is difficult to obtain materials in vitro, the establishment of non-immunogenic neural stem cell lines is difficult, and the difference of species and sociology of allogeneic transplantation. The application of neural stem cells is greatly restricted by ethics and other problems. There are a certain number of bone marrow stromal cells (MSCs) in adult animal bone marrow mesenchymal stem cells (BMSCs) can be transferred to neural stem cells (NSCs) and neurons under appropriate conditions. Some breakthroughs have been made in the research of differentiation into neuron-like cells induced by glial cells and glial cells. However, the neuron-like cells that have been induced to form have the problem of survival time is not long and most of them die after 24 hours. Therefore, how to prolong the life span of MSCs-derived neuron-like cells is very important. On the other hand, although MSCs transplantation has made some progress in the treatment of ischemic brain injury, the percentage of MSCs differentiating into glial cells in vivo is larger and the rate of differentiation into nerve cells is smaller. Therefore, if bone marrow stromal cells are differentiated into neuron-like cells in vitro, cell transplantation will achieve twice the result with half the effort. In this experiment, MSCs were isolated, cultured and purified by cell culture technique in vitro. The viability, growth curve and adherent rate of MSCs were measured. The cultured MSCs were identified by morphological method. The successful bone marrow stromal stem cells were subcultured and expanded in vitro to obtain a certain number of cells. Melatonin (MT) and basic fibroblast growth factor (bFGF) were used to prolong the survival time of neuron-like cells and MSCs derived neuron-like cells. The rat model of focal cerebral ischemia (MCAO) was transplanted by cell transplantation. To observe the distribution and survival of implanted cells, and to investigate the effect of implanted cells on the expression of C-myc Erk protein and its mRNA in MCAO rats.
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R329

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