肿瘤细胞来源胞外体的形态学参数分析  

Analysis of the morphological parameters of tumor cell-derived exosomes

作　　者：葛安兴 傅方洁 贾德功 王雪莲[1] 

机构地区：[1]中国医科大学基础医学院病原生物学教研室,辽宁沈阳110001 [2]中国医科大学临床三系98期

出　　处：《中国病原生物学杂志》2015年第4期321-324,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81472439,No.81101989)

摘　　要：目的获得并分析肿瘤细胞来源胞外体的形态学参数。方法收集喉表皮样癌细胞（Hep-2细胞）培养上清,蔗糖/重水密度梯度超速离心法分离纯化胞外体,透射电镜鉴定胞外体;应用计算机数字图像分析系统测量胞外体的长径、短径、周长、面积,计算其体积、等效直径及圆球度;应用SPSS数据处理软件（19.0版本）对采集到的数据进行统计学分析。结果测量胞外体518个。胞外体长径均值为131.10nm,75.68%的长径在100～150nm之间,95%置信区间128.64～133.54nm;短径均值为88.71nm,65.83%的短径在75～105nm之间,95%置信区间86.83～90.51nm;周长均值为411.49nm,76.25%的周长在320～490nm之间,95%置信区间404.79～418.10nm;面积均值为12237.87nm2,63.71%的面积在8 000～13 000nm2之间,95%置信区间11820.41～12655.24nm2;等效直径均值为61.41nm,94.98%的等效直径在43～80nm之间,95%置信区间60.40～62.32nm;圆球度均值为1.34,63.89%的圆球度在1.12～1.52之间,95%置信区间1.28～1.31。结论利用计算机数字图像分析系统可获得胞外体长径、短径、周长、面积、等效直径、圆球度形态学参数。形态学参数的建立为量化胞外体的大小及胞外体鉴定提供了数字化资料。Objectives To determine and analyze the morphometric parameters of tumor-derived exosomes. Methods Culture supernatants of human epithelial type 2 （Hep-2） ceils were collected, and then exosomes were separated and puri- fied using sucrose/deuteroxide density gradient ultracentrifugation. Exosomes were identified with a transmission electron microscope. The long diameter, short diameter, perimeter, and area of exosomes were measured, and the corresponding volume, equivalent diameter, and sphericity were calculated. SPSS data processing software （ver. 19.0） was used to sta- tistically analyze the collected data. Result In total, 618 exosomes were measured. Tile mean value of the long diame- ter was 131.89 nm. Of the exosomes, 75.68~~ had a long diameter from 100 150 nm, with a 95% confidence interval of 128.64--133.54 nm. The mean value of the short diameter was 88.71 nm. Of the exosomes, 65.83~ had a short diame- ter from 76--105 nm, witha95% confidence interval of 86.83--90.51 nm. The mean value of the perimeter was 411.49 nm. Of the exosomes, 6.25% had a perimeter from 320--490 nm, with a 95% confidence interval of 404.79--418. 10 nm. The mean value of the area was 12237.87 nm2. Of the exosomes, 63.71% had an area from 8 000 13 000 nm2, with a 96% confidence interval of 11 820.41--12 655.24 nm2. The mean value of the equivalent diameter was 61.41 nm. Of the exosomes, 94.98% had an equivalent diameter from 43 80 nm, with a 95% confidence interval of 60.40--62.32 nm. The mean value of the sphericity was 1.34. Of the exosomes, 63.89% had a sphericity from 1.12 1.52, with a 95% confidence interval of 1.28--1.31. Conclusion This is the first time a computer digital imaging system was used to determine the morphometric parameters （e. g. long diameter, short diameter, perimeter, volume, equivalent diameter, and sphericity） of exosomes. Determining morphometric parameters provides digital data to quantify the size of exosomes and identify those exosomes.

关 键 词：胞外体 形态学参数 数字图像分析 

分 类 号：R373.21[医药卫生—病原生物学]