不同诱导条件对体外培养视网膜色素上皮细胞表达血小板源性生长因子A的影响

Effect of different culture conditions on the expression of platelet-derived growth factor-A of cultured human retinal pigment epithelium

目的:血小板源性生长因子由多种细胞产生。为验证视网膜色素上皮细胞是血小板源性生长因子A的分泌细胞,在蛋白水平检测不同诱导条件对人视网膜色素上皮细胞表达血小板源性生长因子A的影响。方法:实验于2007-01/04在吉林大学第一临床医院传染科研究室完成,实验方法符合医学伦理学要求。①实验对象:人视网膜色素上皮细胞株(HRPED407)购自广州中山大学细胞库。②实验方法:分别应用体积分数为0.1的血清、50%巨噬细胞调理液、与单核细胞共培养的方法诱导视网膜色素上皮细胞,24h后撤掉各刺激因素继续培养。③实验评估:通过细胞免疫化学方法检测各种培养条件下血小板源性生长因子A在视网膜色素上皮细胞中的表达情况。结果:细胞免疫化学结果显示,有无诱导因素情况下,视网膜色素上皮细胞中血小板源性生长因子A的表达均呈阳性。结论:有无诱导因素情况下,视网膜色素上皮细胞均分泌血小板源性生长因子A,提示视网膜色素上皮细胞是血小板源性生长因子A的分泌细胞。

AIM： Platelet-denved growth factor （PDGF） is generated by sorts of cells. In this study, the effects of different culture conditions on the expression of PDGF-A of cultured human retinal pigment epithelium （HRPE） were investigated so as to check HRPE is the secretory cell of PDGF-A. METHODS： The experiment was conducted in the Research Laboratory of Infection, First Clinical Hospital of Jilin University from January to April 2007. The experimental procedure was accorded with the Hospital Ethics. （1）HRPE （cell bank of Sun Yat-sen University） was cultured in volume fraction 0.1 serum, 50% macrophage conditioned medium and monocyte as exogenous induce factor, respectively. After 24 hours, the exogenous induce factors were removed. （2）The expression of PDGF-A was detected by immunochemistry. RESULTS： Immunochemistry results showed that all the expressions of PDGF-A were positive in HRPE with or with no induction. CONCLUSION： PDGF-A is secreted by RPE with induction or not, indicating RPE is the secretory cell of PDGF-A.