鼠外周血中转基因胎儿细胞的检测及意义  

Analysis of the status of mouse fetal transgenic cells existed in the maternal peripheral blood

作　　者：颜景斌[1] 王舒[1] 巩芷娟[1] 黄淑帧[1] 

机构地区：[1]上海交通大学医学遗传研究所,200040

出　　处：《中华检验医学杂志》2005年第7期732-735,共4页Chinese Journal of Laboratory Medicine

摘　　要：目的探讨仔鼠出生前后小鼠胎儿细胞在母鼠体内存在的情况,为应用循环于母血中的胎儿细胞进行基因诊断提供科学依据。方法将非转基因的正常母鼠和以HS2、HS3元件、β珠蛋白基因启动子指导的红系特异表达绿色荧光蛋白(GFP)基因的转基因公鼠交配,仔鼠出生前后采集母鼠的外周血用流式细胞仪检测其中表达GFP细胞的比例,并用PCR对母鼠和仔鼠的鼠尾DNA及母鼠的外周血进行检测。结果在仔鼠出生前母体外周血中未检出表达GFP的细胞,而分娩后1～3周此类细胞含量达到峰值,4～5周后将逐步消失。PCR检测证实母鼠体内没有外源GFP基因的DNA片断,但分娩后其外周血中则存在GFP基因阳性片断,这与荧光激活细胞分选仪(FACS)的检测结果一致。结论转基因细胞是一种良好的标记物,可以用来观察胎儿细胞在母体中存在的规律,为使用母体外周血进行基因诊断提供了依据。Objective To study the status of mouse fetal transgenic cells existed in the maternal blood in peri-natal period and provide the information for gene diagnosis using fetal cells circulated in the maternal blood. Methods Mating the female normal mice with male transgenic mice integrated with GFP reporter gene driven by [3-globin promoter and HS2-HS3 elements of LCR. Around the offspring were born, maternal blood was collected and GFP expression level was determined with FACS analysis. Meanwhile ,DNA extracted from the tails of the mothers and their offspring as well as the maternal blood were analyzed by PCR. Results GFP positive fetal cells were not found in maternal blood before offspring were born. 1-3 weeks later, GFP positive fetal cells were detected and this population in maternal peripheral blood reached the peak. 4-5 weeks later, they disappeared gradually. PCR results showed no GFP positive band in the mothers. However, a positive fragment of GFP gene was amplified in maternal blood samples after their offspring were born. This result was in accordance with FACS analysis. Conclusion Transgene cells may be useful markers for the study of status of fetal cells existed in the maternal circulating blood. The results would be beneficial for the gene diagnosis using maternal blood as an alternative resource of fetal cells.

关 键 词：外周血 胎儿细胞 仔鼠 荧光激活细胞分选仪 转基因细胞 

分 类 号：R714.5[医药卫生—妇产科学] R446.11[医药卫生—临床医学]