SD 大鼠视网膜微动脉平滑肌细胞分离和消化方法∗

摘要

Objective:To investigate the isolated and digest approach of retinal microartery smooth muscle cells (RMASMCs)of SD rats.Method:SD rats were sacrificed after taking the eyes,using different digestive en-zymes to digest different times,separate retinal microartery.RMASMCs morphology was observed under an inverted microscope and identified by immunohistochemical staining.Results:Acute dissecting microscope successfully sepa-rate branch retinal artery that retinal arteriolar diameter was 40.23±5.61μm.No.1,2,3 digestive enzymes to digest each 10 to 1 6 minutes,RMASMCs gradually increased,to 1 6 min when the largest number of smooth muscle cells, was 13.40±1.30/low magnification,compared with other digestion time,the difference was statistically significant (P <0.05).Cells were observed under an inverted microscope RMASMCs typical fusiform smooth muscle cell mem-brane integrity,smooth edges;its showedα-action positive expression by immunohistochemical staining.Conclusion:Acute phase separation and digestion method can successfully obtain larger quantities and higher quality RMASMCs.%目的：：探讨分离、消化 SD 大鼠视网膜微动脉平滑肌细胞(RMASMCs)的方法。方法：取正常 SD 大鼠眼球,分离视网膜微动脉,利用不同消化酶消化不同时间后,分离 RMASMCs,在倒置显微镜下观察 RMASMCs 形态,并行免疫组化法鉴定。结果：解剖显微镜下成功急性分离视网膜动脉分支,即视网膜微动脉(直径40.23±5.61μm)。1、2、3号消化酶各消化10-16min,RMASMCs 数量逐步增多,16min 时最多(13.4±1.3个/低倍视野),与其它消化时间相比,差异有统计学意义(P <0.05)。倒置显微镜下观察收获的 RMASMCs 呈长梭形、细胞膜完整、边缘光滑；免疫组化染色鉴定其胞浆呈α-action 阳性表达。结论：采用急性分离、逐步消化的方法能成功获取较多数量和较高质量的 RMASMCs。