CAJ | 학술논문

目的：采用3种方法从大鼠T细胞培养上清中提取纯化exosome ，以获取高质量的exosome。方法分别采用Exo-Quick Precipitation提取法、超滤密度梯度离心法、差速离心法提取 T 细胞培养上清中的exosome。利用透射电镜进行形态学观察，2，2-联喹啉-4，4-二甲酸二钠（BCA ）法进行蛋白定量，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE ）分析蛋白表达的差异，Western blotting检测白细胞介素2（IL-2）的表达。结果3种方法均可提取出exosome；ExoQuick Precipitation 提取法、超滤密度梯度离心法所得样本浓度显著高于差速离心法所得样本浓度（P＜0．05）；SDS-PAGE结果显示3种方法所得样本具有蛋白表达强度的差异；Western blotting显示3种方法所得样本均表达IL-2。结论 ExoQuick Precipitation提取法、超滤密度梯度离心法可获得高纯度、无蛋白丢失的exosome样本。
목적：채용3충방법종대서T세포배양상청중제취순화exosome ，이획취고질량적exosome。방법분별채용Exo-Quick Precipitation제취법、초려밀도제도리심법、차속리심법제취 T 세포배양상청중적exosome。이용투사전경진행형태학관찰，2，2-련규람-4，4-이갑산이납（BCA ）법진행단백정량，십이완기류산납-취병희선알응효전영（SDS-PAGE ）분석단백표체적차이，Western blotting검측백세포개소2（IL-2）적표체。결과3충방법균가제취출exosome；ExoQuick Precipitation 제취법、초려밀도제도리심법소득양본농도현저고우차속리심법소득양본농도（P＜0．05）；SDS-PAGE결과현시3충방법소득양본구유단백표체강도적차이；Western blotting현시3충방법소득양본균표체IL-2。결론 ExoQuick Precipitation제취법、초려밀도제도리심법가획득고순도、무단백주실적exosome양본。
Objective The purification methods of the exosomes derived form T cells were established in order to get high quan-tity exosomes .Methods Exosomes from T cells culture supernatants were purified by ExoQuick Precipitation ,ultrafiltration and sucrose gradient centrifugation ,differential ultracentrifugation ,and confirmed via using transmission electron microscopy .The pro-tein expression of the exosomes were analyzed by SDS-PAGE electrophoresis .Western blotting was used to test the expression of IL-2 .Results The protein concentration of the exosomes purified through ExoQuick Precipitation ,ultrafiltration and sucrose gradi-ent centrifugation were higher than through differential ultracentrifugation (P<0 .05) .SDS-PAGE displayed the difference among the exosome purified by three methods .Three kinds of exosomes all expressed IL-2 .Conclusion ExoQuick Precipitation ,ultrafiltra-tion and sucrose gradient centrifugation technique can obtain high purity and complete exosome sample .