三种常用的密度梯度离心方法富集有核红细胞的比较  

Comparison of three commonly used density gradient centrifugation methods for enrichment of nucleated red blood cells

作　　者：隋玉杰[1] 李凌云[2] 吴飞[2] 杜珍武[1,3] 杨麒巍 张桂珍[1,3] 宋旸[3] SUI Yujie;LI Lingyun;WU Fei;DU Zhenwu;YANG Qiwei;ZHANG Guizhen;SONG Yang(Key Laboratory for Molecular and Chemical Genetics of Critical Human Diseases of Jilin Province,Jilin University Bethune Second Hospital,Changchun 130041,China 2.Department of Gynecology and Obstetrics,Jilin University Bethune Second Changchun 130041,China;Hospital Changehun 130041,Chin;Department of Orthopedics,Jilin University Bethune Second Hospital,Changchun 130041,China)

机构地区：[1]吉林大学第二医院研究中心,长春130041 [2]吉林大学第二医院妇产科,长春130041 [3]吉林大学第二医院骨科,长春130041

出　　处：《中国体视学与图像分析》2018年第3期247-253,共7页Chinese Journal of Stereology and Image Analysis

基　　金：国家自然科学基金青年科学基金项目(81702195);国家发改委建设资金项目(吉大二院精准医学分子诊断应用示范中心建设);吉林省科技厅优秀青年人才基金项目(20180520125JH);吉林省教育厅"十三五"科学技术研究项目(JJKH20180103KJ)

摘　　要：目的探讨富集母血中胎儿有核红细胞的适宜密度梯度离心方法。方法对10例脐血分别用Percoll 1090单密度梯度离法、Percoll 1075与Percoll 1090双密度梯度离心法、Ficoll 1077单密度梯度离心法3种方法进行有核红细胞的分离,然后用适宜的密度梯度离心法分离12例10 mL母血中的胎儿有核红细胞。结果 (1)Percoll 1090单密度梯度离法每mL脐血富集(70. 58±5. 67)×10~4个有核红细胞;Percoll 1075与Percoll 1090双密度梯度离心法每mL脐血富集(23. 62±1. 90)×10~4个有核红细胞; Ficoll1077单密度梯度离心法每mL脐血富集(4. 53±0. 26)×10~4个有核红细胞。(2)采用Percoll 1090单密度梯度离法从12例10 mL母血中富集到的平均有核红细胞数为12. 9±5. 4个。结论 Percoll 1090单密度梯度分离法可有效增加脐血中有核红细胞的分离数量,并从孕妇外周血中成功富集到胎儿有核红细胞。Objective To search for a suitable density cleated red blood cells from maternal blood. Methods gradient centrifugation method to separate fetal nu- Three kinds of common density gradient ccntrifu- gation were used to separate nucleated red blood cells from 10 cases of umbilical cord blood, and then the fetal nucleated red blood cells were separated from 12 cases of 10 mL the maternal blood by the suitable density gradient centrifugation. Results ①（70. 58 ± 5.67） ×10^4nucleated red cells per mL umbilical blood were enriched by Percoll 1090 single density gradient centrifugation; （23.62±1.90） ×10^4 nucle- ated red cells per mL umbilical blood were enriched by Percoll 1075 and Percoll 1090 double density gra- dient centrifugation; （4.53 ± O. 26） ×10^4 nucleated red cells per mL umbilical blood were enriched by Ficoll 1077 single density gradient centrifugation. ②The average number of nucleated red blood cells en- riched from 12 cases of 10 mL maternal blood by Percoll 1090 single density gradient method was 12. 9±5.4. Conclusions Percoll 1090 single density gradient separation method can effectively increase the number of nucleated red blood cells separated from cord blood and successfully enrich fetal nucleated red blood cells from maternal peripheral blood.

关 键 词：密度梯度离心法 脐血 有核红细胞 孕妇外周血 

分 类 号：R715.5[医药卫生—妇产科学]