大鼠骨髓间充质干细胞视网膜下移植的短期观察

目的探索骨髓间充质干细胞（MSCs）视网膜移植的可行性。方法采用贴壁筛选法分离、培养SD大鼠骨髓MSCs，将溴脱氧尿嘧啶（BrdU）标记的大鼠MSCs悬液经玻璃体腔注入大鼠视网膜下腔。术后14d处死大鼠，取新鲜眼球，连续冰冻切片，经BrdU单抗、FITC标记二抗和Rhodopsin单抗、Cy3标记二抗免疫荧光双重标记后，荧光显微镜下观察。结果荧光显微镜下可见移植后的MSCs主要分布于视网膜色素上皮层和视锥、视杆细胞层，未表达视网膜光感受器细胞的特异性抗原视紫红质。结论移植14d后MSCs可在视网膜下腔存活，主要分布于视网膜色素上皮层和视锥、视杆细胞层，未分化成视网膜光感受器细胞。

Short-term observation of transplantation of mesenchymal stem cells into rat sub-retina space

Objective Previous investigations have demonstrated that Mesenchymal stem cells(MSCs)could differentiate into chondrogenic,adipogenic,and osteogenic cell lineages both in vitro and in vivo.This study was designed to transplant BrdU-labeled MSCs into rat retina and observe their survival and differentiation.MethodsMSCs were isolated and cultured by their adherent ability to the wall of culture dishes.Harvested MSCs were characterized by flow cytometry:negative for CD45 and positive for CD90.BrdU-labeled MSCs suspension were injected into the sub-retina space of rat eyes.The rats were sacrificed 14 days after operation.The enucleated fresh eyeballs were frozen and the section was prepared for BrdU monoclonal antibody,FITC-labeled secondary antibody and rhodopsin monoclonal antibody,Cy3-labeled secondary antibody staining.The sections were observed under the fluorescent microscope.ResultsMore than 93% of the 3th passage MSCs showed positive reaction for CD90 and absent reaction for CD45.The nuclear of BrdU-labeled MSCs showed the brown-red staining and unlabeled MSCs were not dyed.The transplanted cells were found mainly in retinal pigment epithelium layer and cones and rods layer,and the engrafted MSCs did not express rhodopsin under fluorescent microscope.ConclusionThe transplanted MSCs could survive in sub-retina space for 14 days.The majority of the cells stay in retinal pigment epithelium layer and cones and rods layer,but they do not differentiate into photoreceptor cells.