免疫磁珠体外纯化人胎脑中CD133阳性干细胞的实验研究

为了探讨免疫磁珠体外纯化人胎儿脑内 CD13 3阳性细胞的可行性 ,本研究取人胎儿脑室下区并制备单细胞悬液 ,采用磁珠分选法选择 CD13 3阳性细胞群 ,台盼蓝染色观察活力并采用流式细胞仪鉴定纯化前后 CD13 3的阳性表达率。结果显示 ,分选后所得细胞纯度为 ( 85 .5 7± 1.66) % ,回收率为 ( 62 .3± 18) % ,纯化前后细胞活力无显著性差异 ( P>0 .0 5 )。结论 :联合应用CD13 3表面标志及免疫磁珠分选系统可有效地从人胎儿脑组织中直接分离得到高纯度的 CD13 3阳性干细胞 ,细胞活力不受影响

PURIFICATION OF CD133 POSITIVE STEM CELLS FROM HUMAN FETAL BRAIN BY USING IMMUNOMAGNETIC BEADS

To determine the possibility and efficiency of purification of CD133 positive stem cells from human fetal brain using immunomagnetic beads, the single cell suspension from subventricular zone in human fetus was prepared and incubated with PE labeled anti-CD133 antibody. The labeled cells were subsequently incubated with anti-PE magnetic beads and separated through magnetic cell sorting system. The viability of cells was evaluated under light microscopy, and the rate of CD133 positive cells in pre- and post-separation groups was assessed by flow cytometry. The results showed that the purity of CD133 positive cells amounted to (85.57±1.66)% after separation, and the mean recovery rate was (62.3±18)%. The viability of cells was not significantly impaired (P>0.05) during the purification process. The study indicates that the highly purified and viable CD133 positive stem cells can be directly isolated from human fetal brain using MACS in combination with CD133 superficial marker.