免疫磁珠法分离白血病转基因小鼠骨髓造血干细胞

本文分离转基因小鼠骨髓造血干细胞。运用针对小鼠干细胞表面特异表达的干细胞抗原 1(stemcellantigen 1,Sca 1)的单克隆抗体和包被于磁颗粒表面的第二抗体 ,采用磁吸附细胞分选方法 (MACS )分离小鼠骨髓造血干细胞 ,用流式细胞术 (FACS )检测MACS分离后骨髓细胞中干细胞 (Sca 1阳性细胞 )比例 ,监测细胞分选效果。结果显示MACS分离后骨髓细胞中Sca 1阳性细胞所占比例达 85 %以上 ,涂片观察发现细胞组成和细胞形态学特征与FACS所得结果一致。MACS可以从转基因小鼠全骨髓细胞中分离出Sca 1阳性的骨髓造血干细胞 ,细胞纯度可达 85 %以上。

Isolation of Bone Marrow Hematopoietic Stem Cells from PML-RARα Transgenic Mice

The bone marrow hematopoietic stem cells were isolated from the PML RARα transgenic mice by means of magnetic absorption cell sorting using two kinds of antibodies:(1)rat anti mouse monoclonal antibody that can specifically bind to the stem cell antigen 1(Sca 1) expressed on the surface of hematopoietic stem cells.(2)Second antibody conjugated to the magnetic beads Flow cytometry was used to monitor efficiency of the isolation method It was found that the proportion of Sca 1 positive cells within the collected cells could be as high as more than 85% after isolation of the hematopoietic stem cells from bone marrow by this method.Cytology examination revealed co incidence with the results obtained by FACS analysis It concludes that magnetic absorption cell sorting method can be used to isolate the hematopoietic stem cells from bone marrow in the transgenic mice and the purity of the isolated cell proportion was more than 85%.