免疫磁珠分选系统在分离大鼠骨髓干细胞群中的应用

采用免疫磁珠分选系统(MACS)分离大鼠骨髓Thy-1.1 干细胞群。收集大鼠胫骨和股骨的骨髓细胞,Percoll密度梯度离心分离单个核细胞,Thy1.1单抗标记,间接MACS分离纯化Thy-1.1 干细胞群,流式细胞仪检测其纯度和分选前后CD34 百分比,台盼兰拒染法检测细胞活力,计算回收率。结果表明分选后的Thy-1.1 细胞纯度达94.2%,回收率64.7%;分选后细胞活力为99.6%,与分选前99.8%无明显差异;分选前CD34 百分比为1.2%,与分选后1.3%无差异。MACS能有效分选大鼠骨髓Thy-1.1 干细胞群,所得细胞纯度高,细胞活力保持好。大鼠Thy-1.1抗原与CD34分子无明显相关性。

Application of Magnetic Activated Cell Sorting in the Isolation of Rat Bone Marrow Stem Cells

To isolate Thy-1.1~ rat bone marrow stem cells by magnetic activated cell sorting(MACS),rat bone marrow cells were collected from rat tibias and femurs,and mononeuclear cells were isolated by Percoll density gradient centrifugation.After incubation with the Thy-(1.1) monoclonal antibody,the Thy-1.1~ stem cells were purified by indirect magnetic activated cells sorting.The cell purity and viability were evaluated by flowcytometry and typanblue staining respectively and the reclaimation rate was calculated.Simultaneously,the percentage of CD34~ cells before and after MACS were detected by flowcytometry.Results showed that the purity of Thy-1.1~ cells was 94.2% after MACS,and the viability was 99.6%,which had no significant difference with the values before MACS(99.8%).The reclaimation rate was(64.7%).The percentage of CD34~ cells before MACS was 1.2%,with no significant difference from the 1.3% after MACS.High purified Thy-1.1~ cells were obtained from rat bone marrow using MACS,and the viability of the cells was not impaired.There was no obvious relationship between the Thy-1.1 antigen and the CD34 molecule of rats.