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目的 观察不同浓度睫状神经营养因子 (ciliary neurotrophic factor,CNTF)对培养大鼠视网膜神经节细胞 (retinal ganglion cell,RGC)生长、存活的影响。 方法 取 15只生后 2～ 3d Wistar大鼠视网膜组织进行细胞培养 ,通过 Thy- 1单克隆抗体免疫细胞化学对培养的 RGC进行鉴定。实验分对照组和10、2 0、40 ng/ ml CNTF组 ( 、 、 组 ) ,记录 RGC存活时间 ,将培养第 3、5、7天的 RGC行四甲基偶氮唑盐 (methylthio- tetrazole,MTT)法测量吸光度 (A)值 [旧称光密度 (OD) ]。 结果 Thy- 1单克隆抗体免疫组织化学检查显示培养 3d的存活细胞 90 %以上为 RGC。细胞存活期间实验组与对照组细胞均无明显突起 ,细胞体积无明显增大 ,实验组细胞存活时间比对照组长 3～ 4d。培养第 5、7d, 组 A值分别为 0 .0 75 8± 0 .0 13 9、0 .0 6 93± 0 .0 113, 组 A值分别为 0 .0 90 2± 0 .0 114、0 .0 82 5± 0 .0 12 5 , 组 A值分别为0 .0 792± 0 .0 13 3、0 .0 6 5 3± 0 .0 0 86 ,对照组 A值分别为 0 .0 6 2 0± 0 .0 0 71、0 .0 5 13± 0 .0 0 6 8。实验组与同时间对照组 A值相比差异有显著性的意义 ( 组与对照组相比 P<0 .0 1, 、 组与对照组相比 P<0 .0 5 )。 结论 一定浓度的 CNTF能促进培养大鼠 RGC的存活 ,?
Objective To observe the effects of different concentrations of ciliary neurotrophic factor (CNTF) on the growth and survival of cultured rat retinal ganglion cells (RGC). Methods Retinal tissues of 15 Wistar rats aged 2 ~ 3 days after birth were cultured in vitro. The cultured RGCs were identified by Thy-1 monoclonal antibody immunocytochemistry. The experimental control group and 10,2 0,40 ng / ml CNTF group (group), the survival time of RGC was recorded, and the RGCs on the 3rd, 5th and 7th day after culture were treated with methylthio-tetrazole , MTT) method to measure absorbance (A) value [formerly known as optical density (OD)]. Results Thy-1 monoclonal antibody immunohistochemical examination showed that more than 90% of surviving cells cultured for 3 days were RGCs. During cell survival, there was no obvious protrusion in the experimental group and the control group, the cell volume did not increase significantly, and the cell survival time in the experimental group was 3 ~ 4 days longer than that in the control group. On the 5th and 7th day of culture, the values of A were 0. 758 ± 0. 0 13 9, 0 6 93 ± 0. 0 113 respectively, and the values of A were 0 90 2 ± 0 0 114, 0. 82 82.5 ± 0 .0 12 5, group A values were 0 .0 792 ± 0 .0 13 3,0 .0 6 5 3 ± 0. 0 0 86, the control group A value was 0 .0 6 2 0 ± 0 .0 0 71,0 .0 5 13 ± 0 .0 0 6 8. There was significant difference between the experimental group and the control group at the same time (P <0.01, P <0.05 compared with the control group, P <0.05). Conclusion A certain concentration of CNTF can promote the survival of cultured rat RGCs.