视网膜细胞培养上清液对骨髓基质细胞的诱导分化作用  

RAT BONE MARROW STROMAL CELLS HAVE POTENTIAL TO DIFFERENTIATE INTO NEURONS,GLIAS AND PHOTORECEPTORS AFTER INDUCED BY SUPERNATANT OF CULTURED RETINAL CELLS

作　　者：万瑾[1] 胡宝洋[1] 刘坤[1] 郑华[1] 肖虹蕾[1] 佘振珏[1] 周国民[1] 

机构地区：[1]复旦大学上海医学院人体解剖学与组织学胚胎学系,上海200032

出　　处：《解剖学报》2006年第3期285-289,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30270699)

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。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 ceiLs 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 supernatunt. 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 supernatunt. In addition only induced BMSCs by supernatunt 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 E17 supernatant. Conclusion BMSCs have potential to differentiate into neurons, glials induced by supematunt of cultured retinal cells. BMSCs induced by supernatant of postnatal retinal cells retinal cells can differentiate into retinal cells.

关 键 词：骨髓基质细胞 视网膜 诱导分化 细胞培养 大鼠 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]