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观察骨髓间充质干细胞(mesenchymal stem cells,MSCS)在体外不诱导的条件下视网膜移植于正常或大鼠Nd:YAG激光损伤视网膜后的定位以及蛋白表达情况.体外培养大鼠MSCS,用荧光染料DAPI标记MSCS,视网膜下移植后分别于10,20,35和50天处死动物,做DAPI荧光观察,并在阳性的连续切片上做神经元核(neuronal nuclei,NeuN)、神经元特异性烯醇化酶(neuronspecific enolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和角蛋白(pallcy-tokeratin,CK)免疫组化染色及HE染色.对损伤大鼠分别于损伤前及损伤后即刻、1-7周检测损伤移植组、损伤组和损伤注射生理盐水组的视网膜电流图(ERG)b波分析.培养的MSCS集落生长迅速,均一性好,DAPI染色观察表明,10天时移植细胞多集中于移植部位周围,正常移植组可见分层,而损伤移植组分布散乱;20天时两组均可见阳性细胞分布范围扩大,分布于视网膜各层;35天正常移植组阳性细胞范围与20天基本相同,而损伤组则进一步扩大,且细胞开始向损伤部位迁移;50天正常移植组荧光范围比35天小,未发现细胞增生,损伤组损伤范围与35天相比扩大不明显,但许多部位有增生.免疫组织化学染色发现阳性区域的细胞在NeuN,NSE,GFAP和CK的表达有不均一性.HE染色表明移植组损伤的恢复好于损伤对照
To observe the localization and protein expression of mesenchymal stem cells (MSCs) in retina after normal or rat Nd: YAG laser-injured retina without inducing in vitro.In vitro MSCs were cultured with fluorescent dye DAPI-labeled MSCs were sacrificed at 10, 20, 35 and 50 days after subretinal transplantation, respectively. DAPI fluorescence imaging was performed. The neuronal nuclei (NeuN) and neuron-specific enolase The neuronspecific enolase (NSE), glial fibrillary acidic protein (GFAP) and pallcy-tokeratin (CK) were detected by immunohistochemistry and HE staining. The retinal current pattern (ERG) b-wave analysis of injury-injured, injured and saline-injected groups was performed immediately, 1-7 weeks after injury.MSCs colonies grew rapidly and with good homogeneity. DAPI staining showed that the number of transplanted cells Mostly concentrated around the transplanted site, the normal graft group can be stratified, while the injury graft group distributed scattered; 20 days when the positive cells were seen in both groups expanded distribution, distributed in the retinal layers; 35 days are positive The number of positive cells in the transplantation group was almost the same as that in the 20-day group, but the damage group was further expanded and the cells started to migrate to the injury site. The fluorescence range of the 50-day normal transplantation group was smaller than that of the 35-day group, and no cell proliferation was observed. Compared with the lesion control group, the number of positive cells in the positive area was significantly higher than that in the neoplastic area (P <0.05). The immunohistochemical staining showed that there was heterogeneity in the expression of NeuN, NSE, GFAP and CK in the positive area