Primary Culture And Biological Characteristics Analysis Of The Transgenic Green Fluorescence Nude Mouse Choroid Plexus Epithelial Cells

Objective : In addition to the secretion of cerebrospinal fluid,the role of choroid plexus epithelial cells, in the central nervous system other effects is poorly understood.The choroid plexus epithelial cells of transgenic green fluorescence nude mouse were cultured in vitro, aimed to develop a kind of natural fluorescent tracing tool of characteristic cells,to provide convenience for further studing.Methods: Pure and flesh choroid plexus tissues were obtained under anatomical microscope, and mechanical separated into cells. Cells were cultured in DMEM medium(F12) containing 20% fetal bovine serum in a culture plate. The proliferation activity were measured by CCK-8 test and Ki-67 staining.TTR, S100, GFAP, alpha-SMA and Nestin immunocytochemical fluorescence staining were performed to identify the cultured cell types. Cell morphology and density were observed under fluorescence microscopeResults: The cultured cells express green fluorescence protein stablely,and emit bright green fluorescence under fluorescence microscope. Cultured cells are polygonal, roomy,and, the cell body is specially big.Almost every cell highly expressed TTR,which is the specific protein of choroid plexus epithelium. In addition, of the cells were negative for GFAP, alpha-SMA and Nestin. The cultured cells have strong activity of proliferation, and at least 7 generations could be passed in vitro.Conclusion : cultured choroid plexus epithelial cells of transgenic green fluorescence nude mice can be successfully subcultured for 7 generations in vitro, and stably express EGFP, this new kind of cells is expected to be further used as a labled tool in the future experiments.