Objective To observe the effects of subretinal transplantation of rat mesenchymal stem cells (rMSCs) on Sodium Iodate (SI)induced retinal degeneration. Methods One hundred and twenty BrownNorway (BN) rats were divided into three groups including SI injection group,rMSCs transplantation group and normal control group, each with 40 rats. The retinal degeneration was induced by caudal vein injection of SI. The retinal pigment epithelium（RPE）and neural retinal were evaluated by ocular fundus photograph, fluorescein fundus angiography (FFA)，electroretinogram (ERG) and histological approach, and TUNEL（terminal deoxynucleotidyl transferasemediated dUTP nick end labeling ）. CMDiIprelabeled primary rMSCs were transplanted into the subretinal space of SIinduced rats. The survival, integration, and differentiation of rMSCs were observed between 14 day to 60 day after the transplantation.Results The rat retinal function was gradually reduced after14 days of SI injection, with a timedependent manner. After the RPE cells were damaged，the outer segments of photoreceptors became disrupted and shortened until karyopyknosis. The nuclear morphology and positive TUNEL labeling indicated that the death of photoreceptor cells was apoptosis. After rMSCs transplantation, CMDiI labeled donor cells were observed to be scattered in the subretinal space and expressed RPE cell markers. Average amplitude of b wave and Ops (oscillation potential) in ERG improved 27.80%,59.38% respectively after rMSCs transplantation.Conclusions Transplanted rMSCs can survive in subretinal space and differentiate into RPE.
目的 比较使用流式细胞仪355 nm和407 nm激光器激发Hochest33342检测细胞凋亡。 方法 通过ATO药物诱导急性早幼粒白血病细胞（NB4）及血清饥饿法诱导人肺癌细胞（NCl-H292）细胞凋亡，取24、48 h时间点收集细胞，进行Hoechst33342-碘化丙啶（PI）双染，分别在配置有两种激光器的流式细胞仪上检测细胞凋亡。 结果 细胞经处理后24 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292细胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。处理后48 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。两种细胞处理后比处理前凋亡率明显升高，但355 nm激光器与407 nm激光器检测的凋亡结果差异不明显（P＞0.05）。 结论 407 nm激光器激发Hoechst33342可检测细胞凋亡。