嗅鞘细胞（OECs）的分离培养及其应用初探

发布时间：2018-05-14 23:42

  本文选题：脊髓损伤 + 嗅鞘细胞　； 参考：《东北师范大学》2008年硕士论文

【摘要】： 脊髓损伤(spinal cord injury, SCI)是临床上常见病多发病,严重的脊髓损伤多发生在青壮年,并且在我国有逐年增加的趋势。脊髓损伤的治疗没有有效的方法。目前治疗的策略主要局限于预防和减少继发损伤及有助于神经恢复或再生的干预调节。实验表明胚胎脊髓移植、细胞(干细胞)移植、转基因细胞移植、基因治疗等使脊髓功能得到了部分恢复,为脊髓损伤的治疗提供了新的治疗方法。尤其是嗅鞘细胞(Ofactory esheathing cell, OECs)移植修复脊髓损伤在实验和临床应用中获得了初步成功,成为有望解决脊髓损伤这一临床世纪难题的重要方法之一。 本实验主要探讨了小鼠OECs的分离培养、纯化增殖和鉴定方法,并初步研究了OECs对神经细胞生长的作用,目的是摸索出一套快速、稳定、高效的OECs的培养方法,为研究OECs在治疗SCI作用提供细胞来源和理论基础。 实验分别采用4种不同的分离方法进行OECs的分离,获得的细胞悬液在同一条件下进行培养,在第12天时对细胞的数量和形态进行比较和观察。分别采用3种不同的纯化方法对OECs进行纯化,在第12天时对细胞的纯度进行比较。将培养的OECs和背根神经节(dorsal root ganglion,DRG)消化制成细胞悬液,以相同比例接种,接种密度为104/ml ,10天后将联合培养的细胞置于倒置显微镜下进行形态学观察。 实验结果显示,采用机械破碎后胰酶消化再经纱网过滤的分离方法分离OECs的效果明显比其它方法好,分离得到OECs数量最多,并且对细胞的形态没有影响;分离培养的OECs成双极或梭形、多突起形和扁圆形油煎蛋形,形态学特征明显;差速贴壁和Ara-c抑制相结合的纯化培养方法兼收了两者的优点,实验证实是一种简单经济而又有实效的OECs培养方法;OECs对被根神经节细胞生长具有促进作用,揭示OECs对SCI的恢复有促进作用,但是还要有进一步的实验来证明。 本实验摸索出了一套成熟OECs的分离培养纯化方法,为研究OECs生物学特征以及应用提供了稳定的细胞来源,同时实验结果还显示OECs能够促进其它神经细胞的生长,这为OECs自体移植治疗SCI提供了理论基础。
[Abstract]:Spinal cord injury (cord injury, SCI) is a common disease in clinic. Serious spinal cord injury (sci) occurs in young adults and has a tendency of increasing year by year in China. There is no effective treatment for spinal cord injury. Current treatment strategies are limited to prevention and reduction of secondary injury and intervention and regulation that contribute to nerve recovery or regeneration. The results show that embryonic spinal cord transplantation, cell (stem cell) transplantation, transgenic cell transplantation and gene therapy have partially restored spinal cord function, which provides a new treatment for spinal cord injury. In particular, the olfactory esheathing cell, OECs) transplantation of olfactory cells for the repair of spinal cord injury has obtained initial success in experimental and clinical application, and has become one of the important methods to solve the problem of spinal cord injury in the century. This experiment mainly discussed the isolation, culture, purification, proliferation and identification of mouse OECs, and preliminarily studied the effect of OECs on the growth of nerve cells. The purpose of this experiment was to find out a set of rapid, stable and efficient OECs culture method. In order to study the role of OECs in the treatment of SCI to provide a cell source and theoretical basis. Four different methods were used to isolate OECs. The cell suspensions were cultured under the same conditions. The number and morphology of the cells were compared and observed on the 12th day. OECs was purified by three different purification methods, and the purity of the cells was compared on the 12th day. The cultured OECs and dorsal root ganglia root ganglion DRGs were digested into cell suspensions and inoculated in the same proportion. After 10 days of inoculation with 104/ml, the co-cultured cells were observed under inverted microscope. The results showed that the separation of OECs by trypsin digestion and warp net filtration after mechanical crushing was better than that of other methods, and the number of OECs was the most, and the morphology of cells was not affected. The OECs isolated and cultured were bipolar or fusiform, multi-protruded and oiled egg shape, and the morphological characteristics were obvious, and the combination of differential adhesion and Ara-c inhibition combined the advantages of the two methods. It is proved that OECs is a simple, economical and effective method of OECs culture, which can promote the growth of root ganglion cells. It is revealed that OECs can promote the recovery of SCI, but further experiments are needed to prove it. A set of methods for isolation, culture and purification of mature OECs were found in this experiment, which provided a stable cell source for studying the biological characteristics and application of OECs. The results also showed that OECs could promote the growth of other nerve cells. This provides a theoretical basis for OECs autologous transplantation in the treatment of SCI.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329;R651.2

【引证文献】

相关硕士学位论文 前1条

1 乔小飞;小鼠嗅鞘细胞的免疫组化和分类鉴定[D];东北师范大学;2010年

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本文编号：1890027