无动物成分器官培养法保存角膜内皮细胞活性的实验研究  

Animal compound free organ culture for corneal endothelial cells activity: An experimental study

作　　者：魏捷[1] 蒋华[1] 

机构地区：[1]济南军区总医院眼科,山东济南250031

出　　处：《实用医药杂志》2011年第8期739-741,共3页Practical Journal of Medicine & Pharmacy

摘　　要：目的探讨无动物成分(ACF)器官培养保存角膜内皮细胞活性方面的效果。方法将Wistar大鼠角膜16只密闭保存于ACF中4周,葡聚糖T500脱水2 d,作为ACF组。以常规含2%胎牛血清(FBS)的MEM基础培养基组作为对照组。保存结束,计数角膜内皮细胞密度(ECD),免疫组织化学染色法和RT-PCR法检测大鼠角膜内皮层中胞质紧密粘连蛋白1(ZO-1)的表达水平,以评估内皮细胞活性和细胞间紧密连接的屏障功能。结果保存结束后,对照组大鼠角膜的ECD值仅为(1 875±162)个/mm2,ACF组则高达(2 143±169)个/mm2,两组间比较差异显著(P=0.006)。角膜冰冻切片显示ZO-1在保存后大鼠内皮层成功表达。RT-PCR检测表明ZO-1mRNA在ACF组的表达水平高于对照组。结论 ACF器官培养法保存角膜可以更好地保持内皮细胞的活性和紧密连接的屏障功能,在无血清器官培养保存角膜方面具有良好的应用前景。Objective To investigate the efficacy of animal compound free （ACF） free organ culture for corneal endothelial cells activity, as ACF group, compared with the conventionally used basic MEM ＋2% fetal bovine serum （FBS）, as controlled group. Method Sixteen wistar rat corneas were cultured in ACF medium for 4 weeks and then dehydrated in 5% dextran TS00 for 48h. After preservation, endothelial cell densities were counted, the immunohistochemistry and semi-quantitative expression of zonula occludens-l（ZO-1） using RT-PCR in rat corneas whose epithelium had been predissected were conducted as well. Result After cultivation, the mean endothelial cells density of the grafts cultured in MEM＋2%FBS controlled group only achieved 1 875±162 cells/mm^2. The higher mean corneal endothelial cell density was found in ACF group （2143±169 cells/mm^2）（p=0.006）. ZO-1 could be seen in corneal endotheliums of frozen sections. ZO-lmRNA expression in rat corneas without epitheliums exhibited the higher level in ACF medium compared with MEM medium. Conclusion Serum-free ACF medium could preserve rat corneal endothelial cells activity and tight junction integrity better than conventional MEM medium with low FBS. ACF showes great superiority and may play an important part in serum free organ culture for corneas in the future.

关 键 词：角膜内皮细胞 器官培养法 无血清培养 

分 类 号：Q813.11[生物学—生物工程]