荧光染料CFSE标记神经细胞的体外研究  

In Vitro Study of Fluorescent Dye CFSE Labeled Nerve Cells

作　　者：邓逸凡 张鹏[1] 王爱璐 陈钰[1] 邓玉林[1] 马宏[1] DENG Yifan;ZHANG Peng;WANG Ailu;CHEN Yu;DENG Yulin;MA Hong(Beijing Institute of Technology,Beijing 100081)

机构地区：[1]北京理工大学,北京100081

出　　处：《生命科学仪器》2021年第6期22-28,共7页Life Science Instruments

基　　金：空间辐射对机体神经损伤机理及药物防护研究。

摘　　要：目的探究荧光染料CFSE对神经母细胞瘤细胞SH-SY5Y标记的理想条件和跟踪时限。方法采用0、1、2、5、10μmol/L的CFSE标记SH-SY5Y细胞,与未标记的U87 MG细胞共培养,在24、48、72h三个时间点分别用流式细胞仪和荧光显微镜检测细胞的阳性率和荧光强度,以此筛选CFSE标记SH-SY5Y细胞的理想条件;并采用MTS法检测各条件下CFSE对于细胞增殖是否有影响。结果终浓度1μmol/L的CFSE是标记SH-SY5Y细胞的理想条件,此条件下SH-SY5Y细胞的荧光强度在72h后仍能与未标记U87 MG细胞有明显区分,且此条件下的CFSE对SH-SY5Y细胞的增殖无影响。结论在体外正常的培养条件下终浓度1μmol/L的CFSE是标记SH-SY5Y细胞的理想条件,在此条件下CFSE能够标记SH-SY5Y细胞长达72h。Objective to explore the ideal condition and tracking time of fluorescent dye CFSE labeling neuroblastoma SH-SY5Y cells. Methods SH-SY5Y cells were labeled with 0,1,2,5,10μmol/L CFSE and co cultured with unlabeled U87 MG cells. The positive rate and fluorescence intensity of SH-SY5Y cells were detected by flow cytometry and fluorescence microscope at 24,48,72 h, respectively, so as to screen the ideal conditions for labeling SH-SY5Y cells with CFSE, and the effect of CFSE on cell proliferation was detected by MTS method. Results CFSE with 1μmol/L final concentration was an ideal condition for labeling SH-SY5Y cells. Under this condition,the fluorescence intensity of SH-SY5Y cells could still be distinguished from that of unlabeled U87 MG cells after 72 h, and CFSE had no effect on the proliferation of SH-SY5Y cells. Conclusion under normal culture conditions in vitro, CFSE with final concentration of 1μmol/L is an ideal condition for labeling SH-SY5Y cells. Under this condition, CFSE can label SH-SY5Y cells for 72 hours.

关 键 词：CFSE SH-SY5Y U87 MG 共培养 流式细胞术 

分 类 号：Q2[生物学—细胞生物学]