不同培养时长的人外周血淋巴细胞G显带染色体制备的优化  

Optimized Preparation of G-banding Chromosomes from Human Peripheral Blood Lymphocytes with Different Culture Duration

作　　者：彭海英[1,2,3,4] 罗清炳 熊进[1,3,4] 张颖 PENG Hai-ying;LUO Qing-bing;XIONG Jin;ZHANG Ying(Reproductive Medicine Center,Renmin Hospital,Hubei University of Medicine;Biomedical Engineering College,Hubei University of Medicine;Hubei Clinical Research Center for Reproductive Medicine;Shiyan Key Laboratory of Reproduction and Genetics,Renmin Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院附属人民医院生殖医学中心 [2]湖北医药学院生物医学工程学院 [3]湖北省生殖医学临床医学研究中心 [4]生殖与遗传十堰市重点实验室·十堰市人民医院,湖北十堰442000

出　　处：《湖北医药学院学报》2024年第6期623-626,631,共5页Journal of Hubei University of Medicine

基　　金：湖北省教育厅科学技术研究计划指导性项目(B2017110);十堰市科技局科学技术研究与开发项目(15Y43)。

摘　　要：目的:探索外周血淋巴细胞经培养不同时长后制备可用于临床诊断的G显带染色体的可行方法。方法:外周血全血接种后常规培养68~72 h为对照组,根据培养时长分24、48、96、120 h组共4个实验组,对接种细胞培养时间、秋水仙素作用浓度、接种细胞浓度共三个因素进行三轮比较实验,比较不同实验组的分裂指数、染色体显带效果,摸索不同培养时长组的最佳实验条件。结果:外周血淋巴细胞培养48 h组接种22滴外周血全血、秋水仙素工作浓度0.14μg/mL组和培养96 h组接种20滴外周血全血、秋水仙素工作浓度0.06μg/mL制片效果与对照组没有显著差异。结论:不同培养时长的外周血淋巴细胞可通过调整接种细胞量和秋水仙素的工作浓度达到常规培养68~72 h的制片和分析效果,应用临床可实现弹性化批量标本处理,极大提高实验室检测效率。Objective To explore the appropriate methods for obtaining G-banding chromosomes from human peripheral blood lymphocytes with different culture duration for clinical diagnosis.Methods Peripheral blood was cultured for 68~72 h as the control group.Four experimental groups were set up according to the culture duration of 24 h,48 h,96 h,and 120 h.Three rounds of comparative experiments were conducted on the factors of culture duration time,colchicine concentration,and cell concentration.The division index and the effect of chromosome G-banding in different experimental groups were compared to find the optimal experimental conditions for groups with different culture duration.Results There was no significant difference between the experimental groups(48 h culture,22 drops of peripheral blood lymphocytes,colchicine concentration of 0.14μg/mL and 96 h culture,20 drops of peripheral blood lymphocytes,colchicine concentration of 0.06μg/mL)and the control group.Conclusion The desirable G-banding after 68~72 h culture and analysis results can be achieved by adjusting cell concentration and colchicine concentration.By this method,specimens can be flexibly processed in batches,which greatly improves the efficiency of laboratory detection in clinical practice.

关 键 词：外周血淋巴细胞 G带制片 细胞培养时长 核型分析 

分 类 号：R440[医药卫生—诊断学]