【摘要】 目的探讨视网膜细胞培养上清液对骨髓基质细胞(BMSCs)的诱导分化作用。方法原代培养胚龄17d(E17)及生后3d(P3)SD大鼠的视网膜细胞,分别收集培养上清液,过滤后与DMEM按2:3混合,用此混合液诱导BMSCs,倒置相差显微镜下观察细胞形态变化,对诱导后的细胞进行神经丝蛋白(NF)、胶质源性纤维酸性蛋白(GFAP)及视网膜特异性标记物进行免疫化学染色,统计分析阳性细胞数;再用RT-PCR进一步检测诱导结果。结果E17和P3细胞上清液均能诱导BMSCs表达NF[(10.35±2.66)%,(10.64±3.94)%]和GFAP[(27.86±7.97)%,(24.38±5.80)%],但只有P3细胞上清液诱导的BMSCs表达视蛋白(opsin),同时RT-PCR也检测到视细胞发育调节转录因子Crx mRNA的表达。结论视网膜细胞培养上清液可以诱导BMSCs向神经元、胶质细胞与视细胞分化。更多还原

【Abstract】 Objective To investigate the effects of supernatant from cultured retinal cells on the differentiation of bone marrow stromal cells(BMSCs) in vitro. Methods Supernatant of cultured embryonic 17 days and postnatal 3 days retinal cells were collected,filtrated and mixed with DMEM at a ratio of 2:3 respectively.BMSCs were induced by two different mixed culture medium,observed under phase contrast microscope,then characterized by immunostained with NF,GFAP and retinal specific antibodies,positive cells were counted and analysed,RT-PCR was used to further confirm the immunocytochemical results. Results BMSCs expressed NF and GFAP after induced by both supernatant.The percentage of NF positive cells were(10.35±2.66)% and(10.64±3.94)% and that of GFAP positive cells were(27.86±7.97)% and(24.38±5.80)% after induced by E17 and P3 supernatant.In addition only induced BMSCs by supernatant from P3 retinal cells were imrnunostained positive in opsin,a specific antibody of photoreceptors;After 3 weeks induction,Crx,a transcription factor of photoreceptor development was detected in BMSCs induced by P3 supernatant by RT-PCR,and not any retinal specific antibody was stained positive in BMSCs induced by El7 supernatant.Conclusion BMSCs have potential to differentiate into neurons,glials induced by supematant of cultured retinal cells.BMSCs induced by supernatant of postnatal retinal cells retinal cells can differentiate into retinal cells.更多还原