侧平底培养试管培养技术在产前诊断中的应用  

Application of flat-sided culture tubes during prenatal diagnosis

作　　者：刘天盛[1] 黄红倩[1] 苏景玉 覃旺尚 费冬梅[1] 欧阳鲁平 黄敏攀 玉晋武[1] 雷亚琴[1] 欧珊[1] 孙惟佳[1] 邱庆明[1] 郑茜 Liu Tiansheng;Huang Hongqian;Su Jiangyu;Qin Wangshang;Fei Dongmei;Ouyang Luping;Huang Minpan;Yu Jinwu;Lei Yaqin;Ou Shan;Sun Weijia;Qiu Qingming;Zheng Qian(Central Laboratory for Genetic and Metabolic Diseases,Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region,Nanning,Guangxi 530003,China;Rare&Inherited Disease Laboratory/NHS North Thames Genomic Laboratory Hub,Great Ormond Street Hospital for Children NHS Foundation Trust,London WC1N 3BH,UK)

机构地区：[1]广西壮族自治区妇幼保健院遗传代谢中心实验室,南宁530003 [2]英国伦敦大奥蒙德街医院,英国伦敦WC1N 3BH

出　　处：《中华医学遗传学杂志》2023年第10期1306-1311,共6页Chinese Journal of Medical Genetics

基　　金：广西壮族自治区卫生健康委员会自筹经费科研课题(Z20200636)。

摘　　要：目的探讨产前诊断使用侧平底培养试管对绒毛和羊水细胞培养、染色体制备的应用价值。方法收集来自2020年1月至2021年5月在广西壮族自治区妇幼保健院遗传门诊就诊进行侵入性产前诊断孕妇的检验样品,其中绒毛样品157例和羊水样品147例。每例样品分别采用1支侧平底培养试管和1个传统细胞培养方瓶进行2种方法的细胞培养,对2种方法检验对侧平底培养试管法与传统细胞培养方瓶法从细胞接种、细胞培养、染色体制备整个实验过程和耗材进行比较。结果绒毛和羊水样本培养的成功率侧平底培养试管法分别为97.45%(153/157)和97.96%(144/147);传统细胞培养方瓶法分别为98.73%(155/157)和98.64%(145/147),统计学分析2种培养方法成功率无显著差异(P>0.05)。绒毛和羊水样本从接种到细胞收获的平均时间侧平底培养试管法分别为8.45 d和9.43 d;传统细胞培养方瓶法分别为9.05 d和9.54 d,绒毛样本使用侧平底培养试管法平均细胞收获天数少于传统细胞培养方瓶法(P<0.001);羊水样本使用2种方法细胞平均收获时间无显著差异(P>0.05)。整个实验过程侧平底培养试管法均在1支侧平底培养试管中进行,无需细胞转移,而传统细胞培养方瓶法需要更换3个容器和2次样本转移。平均每支侧平底培养试管和每个传统细胞培养方瓶使用培养基的总量分别为3.91 mL和6.26 mL。结论使用侧平底培养试管对绒毛和羊水样本进行细胞培养、制备染色体是一种安全、简便、高效、误差少和成本低的染色体制备技术,具有在产前诊断中推广应用的价值。Objective To assess the value of using flat-sided culture tubes for preparing chromosomes through chorionic villi(CV)and amniotic fluid(AF)cell cultures during prenatal diagnosis.Methods From February to March 2020,157 CV samples and 147 AF samples subjected to prenatal diagnosis at the Maternal and Child Health Care Hospital of Guangxi Zhuang Antonomous Region were selected as the study subjects.For each sample,one flat-sided tube and one flask culture were set up by following the standard protocols.The methods were evaluated by comparing the cell growth,experimental process,quality of chromosome preparation and costs.Results The success rates for the culturing of CV and AF samples by the flat-sided culture tube method were 97.45%(153/157)and 97.96%(144/147),respectively.By contrast,the success rates for the conventional flask method were 98.72%(155/157)for CV and 98.64%(145/147)for AF samples.No significant difference was found between the two methods(P>0.05).The average harvest time required by the flat-sided culture tube method was 8.45 days for CV and 9.43 days for AF cultures,whilst the average harvest time for conventional flask method was 9.05 days and 9.54 days,respectively.The flat-sided culture tube method for CV had required significantly shorter average harvest time than the conventional method(P<0.001).No statistical significant difference was found in the average harvest time for AF by the two methods(P>0.05).The conventional culturing method had required three containers with two sample transfers.By contrast,the flat-sided culture tube method was carried out in one tube without any sample transfer.The average total amount of medium used was 3.91 mL for each flat-sided culture tube and 6.26 mL for each conventional flask.Conclusion The flat-sided culture tube method can provide a simple,cost-effective and error-reducing procedure for the CV and AF samples culture during prenatal diagnosis.

关 键 词：侧平底培养试管 细胞培养方瓶 细胞培养 染色体制备 产前诊断 

分 类 号：R714.5[医药卫生—妇产科学]