摘要
目的研究骨髓间充质干细胞(MSCs)在视网膜色素变性(RP)大鼠体内的分化。方法Lewis大鼠腹腔注射3%NaIO3100mg/kg,建立大鼠RP模型,将体外培养的MSCs植入视网膜下腔,用免疫荧光标记法对MSCs进行追踪,并观察术后第1、2、3、4、5周MSCs在该微环境中的分化。结果术后第1周即可见MSCs位于视网膜色素上皮(RPE)层与光感受器细胞层,但全角蛋白(PCK)及Rhodopsin标记阴性,第3周开始可见MSCs在体内表达PCK及Rhodopsin。结论MSCs植入RP模型大鼠视网膜下腔后可存活,主要分布于RPE层和视锥、视杆细胞层,并表达RPE细胞和光感受器细胞的表面标志。
Objective Age-related macular degeneration（AMD） and retinitis pigmentosa are the major eye diseases that cause blindness.The current therapy of transplanting RPE cells,photoreceptor cells or retinal tissue has achieved a lot.Our study was to access the differentiation of rat mesenchymal stem cells（MSCs） in the microenvironment of retinal degeneration. Methods The retinal degeneration models were induced by 3% NaIO3 100 mg/kg intraperitoneal injection in 25 Lewis rats.MSCs from femur and tibia of Lewis rats were isolated,cultivated and passaged in DMEM/F12 containing 10% fetal bovine serum and identified using PE and FITC label.In vitro cultured and BrdU-labaled MSCs suspension（10 μL） was injected into the subretinal space of the right eyes of rats,and equal volume of PBS was injected in the left eyes at a same way.The eyeballs were enucleated in 7,14,21,28 and 35 days after modeling and the tracer and differentiation of MSCs were observed under the fluorescence microscope. Results The destroy of RPE cells and photoreceptors was found in the modeling eyes.Cultured and passaged MSCs showed positive expression for CD90 and absent reaction for CD45.BrdU-labaled MSCs showed the dark red stain in cellular nuclea.The majority of the transplanted MSCs remained in retinal pigment epithelium layer and photoreceptor layer in the first week after modeling.After 3 weeks,BrdU-labaled MSCs presented with the green fluorescence for FITC,and PCK and rhodopsin were expressed in engrafted MSCs,showing the red fluorescence for Cy3.However,the yellow fluorescence was seen in the control eye. Conclusion MSCs can survive mainly in the outer layer of retina in the microenvironment of retinal degeneration and differentiate forward the RPE cell and photoreceptor.
出处
《眼科研究》
CSCD
北大核心
2008年第11期832-835,共4页
Chinese Ophthalmic Research
关键词
骨髓间充质干细胞
NaIO3
视网膜下腔移植
分化
视网膜变性
mesenchymal stem cells
NaIO3
transplantation in subretinal space
differentiation
retinal degeneration
