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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 investigate the presence of fetal cells in the mother rat before and after birth and to provide a scientific basis for genetic diagnosis of fetal cells circulating in maternal blood. Methods Transgenic males with non-transgenic normal males and red gland specific green fluorescent protein (GFP) genes that were directed by HS2, HS3 elements and β-globin gene promoters were mated. The peripheral blood of maternal mice The proportion of GFP-expressing cells was detected by flow cytometry, and the tail DNA of maternal and pups and the peripheral blood of maternal rats were detected by PCR. Results No GFP-expressing cells were detected in the peripheral blood of maternal offspring before birth, but the content of these cells peaked at 1-3 weeks after delivery and gradually disappeared after 4-5 weeks. PCR showed that there was no DNA fragment of exogenous GFP gene in maternal mice, but positive GFP gene was found in peripheral blood after delivery, which was consistent with the results of fluorescence activated cell sorter (FACS). Conclusion Transgenic cells are good markers that can be used to observe the regularity of fetal cells in the maternal body and provide the basis for gene diagnosis using maternal peripheral blood.