本文選題：脊髓損傷 + 嗅鞘細(xì)胞�。� 參考：《東北師范大學(xué)》2008年碩士論文

【摘要】： 脊髓損傷(spinal cord injury, SCI)是臨床上常見病多發(fā)病,嚴(yán)重的脊髓損傷多發(fā)生在青壯年,并且在我國有逐年增加的趨勢。脊髓損傷的治療沒有有效的方法。目前治療的策略主要局限于預(yù)防和減少繼發(fā)損傷及有助于神經(jīng)恢復(fù)或再生的干預(yù)調(diào)節(jié)。實(shí)驗(yàn)表明胚胎脊髓移植、細(xì)胞(干細(xì)胞)移植、轉(zhuǎn)基因細(xì)胞移植、基因治療等使脊髓功能得到了部分恢復(fù),為脊髓損傷的治療提供了新的治療方法。尤其是嗅鞘細(xì)胞(Ofactory esheathing cell, OECs)移植修復(fù)脊髓損傷在實(shí)驗(yàn)和臨床應(yīng)用中獲得了初步成功,成為有望解決脊髓損傷這一臨床世紀(jì)難題的重要方法之一。 本實(shí)驗(yàn)主要探討了小鼠OECs的分離培養(yǎng)、純化增殖和鑒定方法,并初步研究了OECs對神經(jīng)細(xì)胞生長的作用,目的是摸索出一套快速、穩(wěn)定、高效的OECs的培養(yǎng)方法,為研究OECs在治療SCI作用提供細(xì)胞來源和理論基礎(chǔ)。 實(shí)驗(yàn)分別采用4種不同的分離方法進(jìn)行OECs的分離,獲得的細(xì)胞懸液在同一條件下進(jìn)行培養(yǎng),在第12天時對細(xì)胞的數(shù)量和形態(tài)進(jìn)行比較和觀察。分別采用3種不同的純化方法對OECs進(jìn)行純化,在第12天時對細(xì)胞的純度進(jìn)行比較。將培養(yǎng)的OECs和背根神經(jīng)節(jié)(dorsal root ganglion,DRG)消化制成細(xì)胞懸液,以相同比例接種,接種密度為104/ml ,10天后將聯(lián)合培養(yǎng)的細(xì)胞置于倒置顯微鏡下進(jìn)行形態(tài)學(xué)觀察。 實(shí)驗(yàn)結(jié)果顯示,采用機(jī)械破碎后胰酶消化再經(jīng)紗網(wǎng)過濾的分離方法分離OECs的效果明顯比其它方法好,分離得到OECs數(shù)量最多,并且對細(xì)胞的形態(tài)沒有影響;分離培養(yǎng)的OECs成雙極或梭形、多突起形和扁圓形油煎蛋形,形態(tài)學(xué)特征明顯;差速貼壁和Ara-c抑制相結(jié)合的純化培養(yǎng)方法兼收了兩者的優(yōu)點(diǎn),實(shí)驗(yàn)證實(shí)是一種簡單經(jīng)濟(jì)而又有實(shí)效的OECs培養(yǎng)方法;OECs對被根神經(jīng)節(jié)細(xì)胞生長具有促進(jìn)作用,揭示OECs對SCI的恢復(fù)有促進(jìn)作用,但是還要有進(jìn)一步的實(shí)驗(yàn)來證明。 本實(shí)驗(yàn)摸索出了一套成熟OECs的分離培養(yǎng)純化方法,為研究OECs生物學(xué)特征以及應(yīng)用提供了穩(wěn)定的細(xì)胞來源,同時實(shí)驗(yàn)結(jié)果還顯示OECs能夠促進(jìn)其它神經(jīng)細(xì)胞的生長,這為OECs自體移植治療SCI提供了理論基礎(chǔ)。
[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.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R329;R651.2

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 喬小飛;小鼠嗅鞘細(xì)胞的免疫組化和分類鑒定[D];東北師范大學(xué);2010年

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本文編號：1890027