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目的 观察体外培养的牛眼虹膜色素上皮 (iris pigment epithelium,IPE)细胞是否具有与视网膜色素上皮 (retinal pigment epithelium,RPE)细胞相似的特异性吞噬视网膜感光细胞外节 (retinal outersegment,ROS)的功能。 方法 采用 Hu方法分离和培养新生牛眼 IPE细胞。以鼠抗细胞角蛋白抗体和鼠抗 S10 0抗体为一抗 ,采用免疫组织化学染色对培养的 IPE细胞进行鉴定。 Trizol一步法提取生长近融合的传 2代 IPE细胞的总 RNA。根据 Genbank中牛巨噬细胞甘露糖受体的相关系基因列设计特异引物 ,对其 m RNA的表达进行逆转录聚合酶链反应 (reverse transcription polymerase chain- reaction,RT- PCR)检测。 结果 培养获得的 IPE细胞无其它细胞污染 ,提取的 IPE细胞总 RNA完整 ,无降解。RT- PCR检测结果显示 ,体外培养牛眼 IPE细胞可表达甘露糖受体的 m RNA。 结论 体外培养的 IPE细胞可表达甘露糖受体的 m RNA,有可能具有与 RPE细胞相似的特异性 ROS吞噬功能
Objective To observe whether the cultured iris pigment epithelium (IPE) cells have the function of phagocytosis of retinal outersegment (ROS) cells similar to that of retinal pigment epithelium (RPE) . Methods Hu neonatal IPE cells were isolated and cultured. The anti-cytokeratin antibody and the mouse anti-S10 0 antibody were used as primary antibodies, and the cultured IPE cells were identified by immunohistochemical staining. Trizol One-step extraction of total RNA from growing passively fused 2nd passage IPE cells. Specific primers were designed according to the gene sequence of the mannose receptor of bovine macrophage in Genbank, and the reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of m RNA. Results The cultured IPE cells showed no other cell contamination. The total RNA extracted from the IPE cells was intact without degradation. RT-PCR results showed that cultured bovine IPE cells could express m RNA of mannose receptor. Conclusions IPE cells cultured in vitro can express mRNA of mannose receptor and may have specific ROS phagocytosis similar to that of RPE cells