大鼠骨髓间充质干细胞视网膜下移植观察

目的：鉴定骨髓间充质干细胞（mesenchymal stem cells,MSCs)在体外不诱导的条件下视网膜下移植后的定位。方法：体外培养雄性大鼠MSCs，直接作为细胞供体视网膜下移植于成年雄性大鼠视网膜下，4周后将动物处死，取眼球做石蜡切片，用Y染色体鉴定，为做进一步验证，将MSCs用重组腺相关病毒AAV－gfp感染后移植，分别于4、8周取动物眼球作冰冻切片，于荧光显微镜下做绿色荧光蛋白（green fluorescence protein，GFP）表达观察。结果：培养的MSCs集落生长迅速，均一性好；Y染色体原位杂交鉴定表明来源于MSCs的阳性细胞融合入了原来的视网膜结构，在光学显微镜下可分布于视锥，视杆细胞层，双极细胞层及节细胞层；在荧光显微镜下可见GFP标记的阳性细胞存在，分布于视网膜色素上皮层、视锥，视杆细胞层，双极细胞层及节细胞层，细胞形态与结构同周围的视网膜相似；2种标记方法检测到的视网膜结构完整，未见到玫瑰花结样结构。结论：MSCs可在视网膜下移植后4周与原视网膜结构相融合，2种方法检测到的阳性细胞分布于视网膜色素上皮层，视细胞层，双极细胞层及节细胞层。

Observation of rat bone marrow mesenchymal stem cells after subretinally transplanted into rat retina

ObjectiveTo trace bone marrow mesenchymal stem cells (MSCs) after subretinally transplanted into retina without in vitro different induction.MethodsMale rat MSCs were cultured and were either subretinally transplanted into rat eye or transduced with recombinant adeno-associated virus cloned with green fluorescence protein (GFP) beforehand. Four and 8 weeks later Cross-sex transplantation and GFP detection were used to trace the origin of the repopulating retina cells. Markers for Y chromosome and GFP fluorescence are used to identify retina cells of bone marrow origin.ResultsThe positive cells are found in retinal pigment epithelium(RPE) layer, photoreceptor layer, bipolar layers and strata ganglionare. No formation of rosettes was found at the end of the observation.ConclusionFrom these cells, a proportion of the transplanted MSCs can interfuse into retina in vivo and the positive cells are found in RPE layer, inner and outer nuclear layers and ganglion layer.