Tf-USPIO纳米微粒的合成及TfR转基因神经干细胞过表达的研究

Study on the synthesis of Tf-USPIO nanometer particle and overexpression of TfR transgenic neural stem cells

目的合成Tf-USPIO纳米微粒以及建立稳定表达TfR的神经干细胞。方法利用慢病毒转染神经干细胞,通过流式细胞仪筛选出稳定表达TfR的神经干细胞,运用WB检测其表达水平。将Tf与USPIO进行偶联,合成Tf-USPIO纳米微粒,通过DLS方法测出USPIO和Tf-USPIO的直径分别为(36±0.7)nm,(43.5±0.08)nm。结果慢病毒成功的转染神经干细胞,通过流式细胞仪器筛选出稳定表达的神经干细胞,并用WB证实了TfR的过表达;通过偶联的方法合成了稳定的Tf-USPIO微粒。结论成功构建了TfR神经干细胞系,合成出稳定的Tf-USPIO纳米微粒。

Objective To synthesize Tf-USPIO nanometer particle and establish the neural stem cells system that can express transferrin receptors （ TfR） stably.Methods The neural stem cells were transfected by lentivirus,then the neural stem cells that could express TfR stably were selected out by flow cytometry.The expression levels of TfR were detected by Western Blotting.The TF was coupled with USPIO to synthesize Tf-USPIO nanometer particle,which were measured by DLS method,with the diameters of USPIO and Tf-USPIO being （36 ±0.7） nm,（43.5 ±0.08） nm, respectively.Results The neural stem cells were transfected by lentivirus successfully, and the neural stem cells that could express TfR stably were selected out by flow cytometry successfully,moreover, the overexpression of TfR was identified by Western Blotting.The stable Tf-USPIO nanometer particles were synthesized by link-coupled method.Conclusion The TfR neural stem cells line is successfully established,which can synthesize stable Tf-USPIO nanometer particles.