睾丸支持细胞对体外培养鼠胎脑皮质细胞的影响.docVIP

睾丸支持细胞对脑皮质细胞的 赵 佳，金 洁，梁元晶，史小林 Effects of Sertoli cells on mouse embryonic cerebral cortical cells in vitro Zhao Jia, Jin Jie, Liang Yuan-jing, Shi Xiao-lin Abstract BACKGROUND: Functioning cell is focus of current studies to prove some growth factors for improving survival environment of cells. Sertoli cells can secrete plenty of nutritive substances and growth factors, and its Fas ligand can reduce host immunologic rejection following implantation. OBJECTIVE: To investigate the effect of Sertoli cells on mouse embryonic cerebral cortical cells in vitro. DESIGN, TIME AND SETTING: In vitro observation of cytology was performed at the laboratory of Reproductive Medical Center, Capital Medical University between October 2005 and September 2007. MATERIALS: Forty SPF male ICR mice, aged 10-15 days, were selected to harvest Sertoli cells; forty 15-day-old fetal mice were selected to harvest cerebral cortical cells. METHODS: The density of isolated cerebral cortical cells was adjusted to 2×109 L-1 and divided into two groups. The cells in the experimental group were seeded on a culture flask coated with 70% passaged Sertoli cells and added DMEM/F12 culture medium containing 0.1 volume fraction of fetal bovine serum, 1×105 U/L penicillin, and 0.01 g/L streptomycin and incubated in 5% CO2 at 37 ℃ for 17 days. The cells in control group were cultured in a flask coated with 0.05% polyadenylic acid. MAIN OUTCOME MEASURES: Cells were identified using immunohistochemistry, immunocytochemistry and immunocytological fluorescent methods. The number of neurons and neurospheres were counted. Tyrosine hydroxylase (TH) positive rate was detected. RESULTS: All cells in two groups were neuron specific enolase, nestin, and TH positive. Compared with 2 hours of culture, the number of attached neurons significantly decreased with time (P 0.01), but the number of attached neurons in the experimental group was still significantly higher than the control group at each time point (P 0.