大肠癌细胞及其上清液体外培养泡球蚴的实验研究  

Experimental studies on the in vitro culture of Echinococcus multilocularis metacestodes with Caco-2 cells and its supernatant fluid

作　　者：马海龙[1] 郭倩[1] 孙世安[1] 李兆勇[1] 

机构地区：[1]武警新疆总队医院,新疆乌鲁木齐830091

出　　处：《武警后勤学院学报（医学版）》2012年第12期998-1000,F0003,共4页Journal of Logistics University of PAP（Medical Sciences）

摘　　要：【目的】利用大肠癌细胞(Caco-2)及其培养上述细胞的上清液,在体外对泡球蚴进行培养,建立稳定的体外培养模型,观察泡球蚴的在体外的生长、发育过程。【方法】培养大肠癌细胞(Caco-2),留取细胞上清液。取泡球蚴组织,分别与大肠癌细胞及其细胞培养上清的DMEM完全培养基于37℃5%CO2培养箱中培养38d。实验期间对囊泡进行计数,观察囊泡生长情况,记录囊泡最大、最小直径,绘制生长曲线。培养结束后,取培养囊泡囊液进行蛋白定量。【结果】泡球蚴与大肠癌细胞及其上清液中均能共同生长,呈出芽生殖方式,囊泡直径0.5～5.0mm;囊液蛋白含量分别为:大肠癌细胞,3.2mg/ml;大肠癌细胞上清液,1.2mg/ml。泡球蚴在体外培养初期呈较快速性生长(1～15d);生长中期可见一平台期(15～26d);后期表现为逐渐减少。【结论】多房棘球蚴体外生长因素不依赖培养细胞本身,利用大肠癌细胞上清液建立多房棘球蚴体外培养模型具有一定的可行性,由于大肠癌细胞(Caco-2)与其培养细胞的上清液所含成分可能不同,可能引起囊泡内囊液的蛋白定量有所差异。[Objective] To establish a model for the in vitro culture of Echinococcus multilocularis metacestodes, and observe the process of growth. [Methods]After recovery Caco-2 cell,and its supernatant fluid,metacestode tissue was placed in DMEM medium containing the cells and its supernatant fluid and cultured in an incubator containing 5% CO2 at 37℃ for a period of 38 days. Number of vesicles was counted,condition of growth was recorded maximum and minimum diameter of vesicle was measured growth curve of secondary vesicles was drawed. Protien level of hydatid fluid of secondary vesicles was determined. [Results]Echinococcus multilocularis metacestodes could develop in DMEM medium containing Caco-2 cell,and its supernatant fluid in vitro. Budding of new cysts were observed and the vesicles’ diameter was from 0.5 to 5mm. Protein level of secondary vesicles＇ hydatid fluid in vitro were determined in Caco-2 cell was 3.2 mg/ml in Caco-2 cells＇ supernatant fluid was 1.2 mg/ml. [Conclusion]Echinococcus multilocularis metacestodes growth in vitro not rely only on Caco-2 cells completely. Improvement in vitro culture model of Echinococcus multilocularis metacestodes was feasible by utilizing host cells＇ supernatant fluid. Different composition of Caco-2 cells and its supernatant resulted in different of protein level secondary vesicles’ hydatid fluid.

关 键 词：泡球坳 体外培养 大肠癌细胞 上清液 

分 类 号：R37[医药卫生—病原生物学]