【摘要】 目的 :利用流式细胞仪研究孕妇血中胎儿细胞对三种抗体表达的情况。希望能更准确的标记孕妇血中的胎儿细胞 ,以助提高胎儿细胞的分离纯度。方法 :用CD71、CD45、GPA(糖蛋白A)分别标记抗凝的 2 2例孕妇血、2 8例脐血、30例正常未孕妇女血各 10 0 μl。在流式细胞仪上检测。同时对已标记 2种抗体的 2例 2 0ml母血在流式细胞仪上进行胎儿细胞分离 ,并经PCR技术鉴定男性SRY基因。结果 :1、正常未孕妇女血中几乎无CD71+细胞存在。 2、在脐血和孕妇血中CD71+细胞主要有两群。高表达群和低表达群。有核红细胞 (表达CD71+GPA + /CD45 - )主要在高表达群中。脐血是 5 8.73% ,孕妇血 36 .43 %。 3、无论是高表达群和低表达群 ,CD45 +细胞占多数。在脐血中低表达群是 90 .86 % ,高表达群占 2 2 .2 6 %。孕妇血CD71+低表达群中CD45 +细胞占 82 .11%。高表达群CD45 +细胞占 79.2 9%。 4、利用流式细胞仪分离已标记三种抗体的母血中胎儿细胞 ,经PCR技术鉴定分离后的标本中存在男性SRY基因的胎儿细胞。结论 :1.正常未孕妇女血中几乎不存在有核红细胞。 2 在脐血和孕妇血中有核红细胞主要有两群。高表达群和低表达群。有核红细胞主要在高表达群中。 3、无论高表达群或低表达群 ,CD45 +细胞 (增殖的白细胞 )仍占多数 ,说更多还原

【Abstract】 Using flow cytometry to assay three antibodies’ (CD71,CD45,Glycophori A GPA)expressions in the fetal nucleated erythrocytes from maternal blood.Methods:100μl anticoagulative maternal blood,cord blood and normal no-pregancy women blood were mixed by these antibodies (CD71-FITC,CD45-Cychrom.Glycophori A-PE GPA)in room temperature respectively.These mixers were performed using non-washing erythrocyte fragmentation buffer.These samples then were assayed in flow cytometry.The results were recorded in computer.At the same time.we isolated these marked male fetal cells from 20ml anticoagulative maternal blood in flow cytometry,These isolated fetal cells were used to confirm fetal sexes by looking for SRY gene sequence in using PCR technique.Results:1. There wasn’t any nucleated erythrocyte in the normal no-pregnancy women blood.2. In the maternal blood and the cord blood,there were two groups nucleated erythrocytes (CD71+,GPA+/CD45-),the antibody higher expression group and lowe expression group.But the numbers of nucleated erythrocytes in higher expression group were more than lower expression group. The cord blood was 58.73%,the maternal blood 36.43%.3.In both groups,the proliferatijng hamoleukocytes(CD71-GPA-/CD45+) still were multiple.4. The male fetus were confirmed in sorted cells from 2 wonem with male babies by using polymerase chain reaction (PCR) to proliferate Y-special repeated sequences (SRY).Conclusing 1.There wasn’t any nucleated erythrocyte in the normal no-pregancy women blood.2.In the pregnant women blood and the cord blood,there were more nucleated erythrocytes in higher expression group than in lower expession group.3.In the pregnant women blood and the cord blood, the proliferating hamoleucocytes still were multiple.So it is a fact that we still lack specific marker to recognize the fetal cells in maternal circulation.CD45 negative choice was recommended.更多还原