骨髓基质干细胞向神经细胞诱导分化及体内移植治疗脑缺血的实验研究

发布时间：2018-06-28 11:43

本文选题：骨髓基质干细胞 + 褪黑素　；参考：《中国医科大学》2005年博士论文

【摘要】：目的神经干细胞的发现及其分离鉴定、培养技术的建立,使神经细胞不可再生的传统理论面临巨大挑战,并为神经系统疾病的治疗提供了新的思路。但神经干细胞数量少、分布分散,体外培养时取材困难,无免疫原性神经干细胞系建立困难,以及异体移植的种属差异和社会学、伦理学等方面的问题都极大地限制了神经干细胞的应用。成体动物的骨髓中存在一定数量的骨髓基质干细胞(MSCs),骨髓基质干细胞在适当条件下可以向神经干细胞、神经元、和神经胶质细胞方向分化。MSCs诱导分化为神经元样细胞的研究工作已有一定突破,但诱导形成的神经元样细胞存在着存活时间不长,24小时后大多死亡的问题。故如何延长MSCs源性的神经元样细胞的寿命是极为重要的。另一方面MSCs移植治疗缺血性脑损伤虽己取得一定的进展,但MSCs移植后在体内环境下分化为神经胶质细胞的比率较大而分化为神经细胞的比率较小,所以如将骨髓基质干细胞在体外分化为神经元样细胞,再行细胞移植将会有事半功倍的效果。本实验应用体外细胞培养技术将大鼠骨髓基质干细胞分离培养和纯化,测定MSCs的细胞活力、生长曲线和贴壁率,用形态学方法鉴定培养的骨髓基质干细胞。将培养成功的骨髓基质干细胞在体外传代、扩增,以得到一定数量的细胞。以褪黑素(MT)和碱性成纤维细胞生长因子(bFGF)予以干预,以期延长诱导的神经元样细胞的存活时间,并应用生存时间较长的MSCs源性神经元样细胞,对局灶性脑缺血大鼠模型(MCAO)进行细胞移植。观察植入细胞的分布及存活情况,并探讨细胞植入后对MCAO大鼠脑神经元C-myc、Erk蛋白及其mRNA表达的影响。
[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.
【学位授予单位】：中国医科大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329

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