基于磁性纳米粒子-抗体复合物分离CD34^+细胞的新技术

A New Method for Isolating CD34^+ Cells Based on Complex of Magnetic Nanoparticles and Antibody

本研究合成磁性纳米粒子-抗体复合物(magnetic nanoparticles-antibody,MNPs-Ab),建立用此复合物从脐带血中分离CD34+细胞的方法并评价其效果。利用磁性纳米粒子-抗体复合物的超顺磁性和对CD34+细胞的识别及结合功能,在外加磁场的作用下,分离脐血中的CD34+细胞;用扫描电子显微镜(scanning electron microscopy,SEM)观察分离后CD34+细胞的形态变化;用流式细胞仪(flow cytometer,FCM)分析分离效率;用液体培养和集落培养鉴定CD34+细胞增殖能力和多向分化能力。结果显示:利用磁性纳米粒子-抗体复合物可以快速、高效地将CD34+细胞从脐血中分选出来,而且分离后的CD34+细胞仍然保持正常形态、高度的增殖能力和多向分化能力。结论:成功地研制了磁性纳米粒子-抗体复合物分离CD34+细胞技术,该技术能高效、快速地分离CD34+细胞。

The purpose of this study was to synthesize the complex of magnetic nanoparticles and antibody, and to apply them to isolate the CD34^＋ cells from umbilical cord blood, then to evaluate its separation efficiency. The complex of magnetic nanoparticles and antibody was used to separate CD34^＋ cells from umbilical cord blood in the outer magnetic field because of its superparamagnetism, specific identification and function of combination with CD34^＋ cells. Scanning electron microscopy was used to observe the morphology of the separated CD34^＋ cells. Flow Cytometer was applied to evaluate the sorting efficiency of magnetic nanoparticles, liquid culture and colony culture were taken to assay proliferation and differentiation capacity of the separated CD34^＋ cells. The results showed that the CD34^＋ cells from umbilical cord blood were isolated fast and effectively by the complex of magnetic nanoparticles and monoclonal antibody. Moreover, the isolated CD34^＋ cells still maintained its normal morphology, highly proliferative and differentiative capacity. It is concluded that the complex of magnetic nanoparticles and monoclonal antibody has been successfully synthesized and developed as a technique which efficiently and quickly isolates CD34^＋ cells from umbilical cord blood.