本文選題：膠質(zhì)瘤 + exosome�。� 參考：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:體外培養(yǎng)U251細胞,分別利用差速離心法和試劑盒兩種方法分離、提純exosome,證明分離提純的物質(zhì)為exosome,并對兩種方法進行優(yōu)缺點比較。方法:體外培養(yǎng)U251細胞,觀察細胞貼壁生長良好后,將培養(yǎng)基更換為不添加小牛血清的培養(yǎng)基,1周后將細胞培養(yǎng)上清液分兩組進行收集,分別以差速離心法、試劑盒提純exosome。然后在電子顯微鏡下通過液相載網(wǎng)法觀察形態(tài),并照相,確認(rèn)exosome存在,對比兩組的形態(tài)結(jié)構(gòu),記錄直徑大小,并對兩種方法進行比較。結(jié)果:U251細胞可以分泌exosome,exosome小囊泡均能夠在差速離心法和試劑盒兩種方法下提取到。通過差速離心機進行差速離心提取的exosome與試劑盒提取的exosome結(jié)構(gòu)相同,電鏡結(jié)構(gòu)為小囊泡狀,表現(xiàn)為球體、橢球體,中央為低密度區(qū),周圍深染。兩組測量直徑均在35~160nm之間,差速離心法組測量直徑均值為99.33±4.21nm,試劑盒組測量直徑均值為98.08±4.13nm,經(jīng)統(tǒng)計學(xué)分析兩組方法提取的exosome直徑隨機抽樣結(jié)果無顯著性差異。結(jié)論:膠質(zhì)瘤細胞可以分泌exosome。差速離心法、試劑盒均可以分離和提純到形態(tài)結(jié)構(gòu)相同的exosome。試劑盒具有操作相對簡單,耗時短,樣本需求量少等優(yōu)點,但提取純度低,科研成本高,臨床難以普遍應(yīng)用。差速離心法操作步驟繁多,耗時長,樣本需求量大,但技術(shù)易掌握,科研成本低,提取純度高,有助于exosome進一步研究以及向臨床應(yīng)用轉(zhuǎn)化。
[Abstract]:Aim: to culture U251 cells in vitro and to isolate and purify U251 cells by differential centrifugation and kit respectively, and compare the advantages and disadvantages of the two methods. Methods: U251 cells were cultured in vitro. After the cells grew well, the culture medium was replaced with the medium without calf serum for 1 week. The supernatant of cell culture was collected into two groups, and the exosome was purified by differential centrifugation and kit. Then the morphology was observed by liquid phase netting method under electron microscope, and the existence of exosome was confirmed. The morphological structure and diameter of the two groups were compared, and the two methods were compared. Results the exosome-exosome vesicles could be obtained by differential centrifugation and kit. The structure of exosome extracted by differential centrifuge was the same as that of exosome extracted by reagent kit. The structure of electron microscope was vesicular, which was sphere, ellipsoid, low density area in the center and deep staining around it. The mean diameter of the two groups was 99.33 鹵4.21 nm in the differential centrifugation group and 98.08 鹵4.13 nm in the kit group. There was no significant difference in the random sampling results of the exosome diameter obtained by statistical analysis between the two groups. Conclusion: glioma cells can secrete exosome. Differential centrifugation and kit can be separated and purified to exosome with the same morphology and structure. The kit has the advantages of relatively simple operation, short time consuming and less sample demand, but the purity of extraction is low, the cost of scientific research is high, and it is difficult to be widely used in clinic. The differential centrifugation method has many steps, long time consuming, large sample demand, but the technology is easy to master, the cost of scientific research is low, and the purity of extraction is high, which is helpful for further study of exosome and its transformation to clinical application.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.41

【參考文獻】

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