培养人视网膜色素上皮细胞中β-半乳糖苷酶活性研究

目的　观察培养人视网膜色素上皮 (RPE)细胞中复制衰老相关的 β 半乳糖苷酶活性。 方法　在培养的RPE细胞中 ,以传代次数 ( 5、10、2 0 )和取材供体的年龄 ( 2 6、44、5 7岁 )分组 ,进行标准化的 β 半乳糖苷酶活性和细胞增殖能力检测。结果　培养RPE细胞中 β 半乳糖苷酶活性阳性细胞数量随传代次数的增加而增加 ,在 2 0代的细胞中 ,阳性细胞的数量为 87%± 1 2 % ,明显高于第 5代和第 10代细胞 (P <0 0 5 )。而细胞的增殖能力明显下降 (P <0 0 5 )。结论　培养RPE细胞复制衰老的发生与细胞传代次数和取材年龄相关 ,其相关研究可能对了解年龄相关性视网膜疾病的发病机制有帮助

Detection of replicative senescence-related beta-galactosidase activity in cultured human retinal pigment epithelium cell

Objective To observe the replicative senescence-related beta-galactosidase activity in cultured human retinal pigment epithelium (RPE) cells. Methods In a series of cultured RPE cells,a standard beta-galactosidase histochemistry technique and MTT assay were performed to detect the activity of beta-galactosidase on a varying number (5,10,20) of population doublings and age (26,44,57) of the (donator. Results) Accumulated beta-galactosidase positive cells in RPE cultures showed a positive correlation to population doublings and donor age(P<0.05).RPE cells proliferation was markedly suppressed (P<0.05). Conclusion The occurrence of cellular replicative senescence in cultured RPE cells accumulate with the increased passages and the age of the donator,which implies that this procedure is associated with intraocular aging related diseases.Similar study may help to answer questions involved in the pathogenesis of age-related retinal disease.