摘要
目的比较新型(卡扣式)载玻片培养盒原位收获法及常规塑料培养瓶消化收获法在羊水细胞染色体核型分析中的应用。方法收集2019年1~12月于我院产科进行羊膜腔穿刺的1568例孕妇的羊水样本,分别采用新型(卡扣式)载玻片培养盒以及常规塑料培养瓶进行羊水细胞培养,细胞收获后按照标准流程进行分带染色及阅片。比较两种羊水细胞培养方法在不同孕周的培养成功率,并对所获得的染色体核型进行计数及分析。结果1568例羊水样本双线培养及制片,其中常规塑料培养瓶法在孕16~17^(+6)周组和孕18~20^(+6)周组各有1例细菌污染,成功率为99.87%(1566/1568)。新型(卡扣式)载玻片培养盒法在孕18~20^(+6)周组有3例细菌污染,有2例因卡扣式培养盒密封处理不合格导致漏液,培养成功率为99.68%(1563/1568)。在不同孕周组,两种方法的培养成功率无显著差异(P>0.05)。新型(卡扣式)载玻片盒培养法和常规塑料培养瓶法所需细胞量分别为6~10 ml和9~12 ml羊水;培养基用量分别为7 ml和9 ml;收获步骤所需时间分别为(105±6)min和(120±10)min。两种培养方法收获制片处理后所获得的平均有效核型数目分别为11个和31个核型。核型分析共发现异常核型98例,包括30例结构异常和68例数目异常,其中染色体嵌合8例。结论两种方法培养制片均可用于羊水细胞培养及核型分析。新型(卡扣式)培养盒原位培养法所需细胞量及培养基用量均少于常规塑料培养瓶法,与之有相同的培养效果。同时使用两种方法可提高异常核型检出率,并可有效解决染色体核型分析中的嵌合问题。
Objective:To compare the new method of in situ culture in slide culture box and the conventional method of digesting in plastic culture bottle for karyotype analysis of amniocytes.Methods:Amniotic fluid samples were collected from 1568 pregnant women who underwent amniocentesis in obstetrics department of our hospital from January to December in 2019.The amniocytes were cultured by new slide culture box method and conventional plastic culture bottle methods respectively.The cells were harvested,stained and film reading according to the standard procedure.The success rates of the two amniocytes culture methods at different gestational weeks were compared,and the karyotypes obtained were analyzed.Results:A total of 1568 amniotic fluid samples were double cultured,and film production were made.One case of bacterial contamination occurred in 16-17^(+6) gestational weeks group and 18 to 20^(+6) gestational weeks group in conventional plastic culture bottle method respectively,with the success rate of 99.87%(1566/1568).Three cases of bacterial contamination occurred at 18 to 20^(+6) gestational weeks in slide culture box method.Two cases of contamination were caused by leakage due to unqualified sealing of slide culture box.The success rate of cultivation was 99.68%(1563/1568).There was no significant difference in the success rate between the two culture methods in different gestational age groups(P>0.05).The slide culture box method and the conventional plastic culture bottle method required 6-10 ml and 9-12 ml of amniotic fluid and 7 ml and 9 ml of the medium amount for amniotic fluid respectively.The harvesting steps required(105±6)min.and(120±10)min.respectively.The average number of effective karyotypes obtained by the two methods was 11 and 31 respectively.Karyotype analysis revealed 98 abnormal karyotypes,including 30 structural abnormalities and 68 number abnormalities,among which 8 were chromosomal chimerism.Conclusions:Both methods can be used for amniocytes culture and karyotype analysis.The amount of cells and medium required by the new method were less than those of the conventional plastic bottle method,and the new method had the same culture effect as the conventional method.Using the two methods at the same time can improve the detection rate of abnormal karyotypes and effectively solve the problem of chimerism in chromosome karyotype analysis.
作者
郝娜
田晓彤
李萌萌
常家祯
周京
戚庆炜
蒋宇林
周希亚
刘俊涛
HAO Na;TIAN Xiao-tong;LI Meng-meng;CHANG Jia-zhen;ZHOU Jing;QI Qing-wei;JIANG Yu-lin;ZHOU Xi-ya;LIU Jun-tao(Department of Obstetrics & Gynecology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,National Clinical Research Center for Obstetrics & Gynecologic Diseases,Beijing 100730;State Key Laboratory of Complex Severe & Rare Diseases,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730)
出处
《生殖医学杂志》
CAS
2021年第6期793-797,共5页
Journal of Reproductive Medicine
基金
生殖健康及重大出生缺陷防控研究(2018YFC1002704)。
关键词
染色体
羊水细胞
原位培养
产前诊断
核型分析
Chromosome
Amniocytes
In situ culture
Prenatal diagnosis
Karyotype analysis
