密度梯度离心和免疫磁珠分选相结合分离脐血中胎儿有核红细胞

Density Gradient Centrifugation and Magnetic Activated Cell Sorting Combined Separation of Fetal Nucleated Red Blood Cells from Cord Blood

目的研究密度梯度离心和免疫磁珠分选相结合方法富集脐血中胎儿有核红细胞。方法对5例脐血首先用密度梯度离心和免疫磁珠分选依次进行胎儿有核红细胞分离,用流式细胞检测和细胞染色对分选前后细胞进行分选纯度分析。结果 1.通过流式细胞仪检测发现CD71单克隆抗体磁珠用于脐血中胎儿有核红细胞分选(CD71+,CD45-),分选后的纯度可以达到81%,而分选前CD71阳性细胞只占23.2%。2.分选前脐血涂片可见大量的成熟红细胞,含少量有核红细胞;密度梯度离心后可见绝大多数细胞为单个核细胞(淋巴细胞、单核细胞和有核红细胞),含少量红细胞;CD71单克隆抗体磁珠分选后阳性细胞多为胞浆丰富略嗜碱性的有核红细胞。结论密度梯度离心和免疫磁珠分选相结合可以有效提高脐血中胎儿有核红细胞的分选纯度。

Objective To investigate Density Gradient Centrifugation and Magnetic Activated Cell Sorting combined separation of fetal nucleated red blood cells(FNRBCs) from cord blood. Methods 5 cases of cord blood FNRBCs separated by Density Gradient Centrifugation and Magnetic Activated Cell Sorting in turn. Using Fluorescence Activated Cell Sorting (FACS) technique and cell staining to conducted purity analysis.Results 1. FACS detectes the purity of CD71+ FNRBCs from cord blood separated by CD71 monoclonal antibody beads can reach to 81% while only 23.2% before the election. 2. Pre-sorting of cord blood smear shows a large number of mature red blood cells and contains little FNRBCs.After Density gradient centrifugation,most cells were mononuclear cells included lymphocytes,monocytes and FNRBCs and little red cells.CD71+ cells separated by CD71 monoclonal antibody beads are most FNRBCs which have basophilic cytoplasm.Conclusion Density Gradient Centrifugation and Magnetic Activated Cell Sorting combined separation can increase the sorting purity of FNRBCs from cord blood.