Sakuragi M, Tomita H, Abe T, Tamai M
Department of Ophthalmology, Tohoku University, School of Medicine, Sendai, Miyagi, Japan.
Curr Eye Res. 2001 Sep;23(3):185-91. doi: 10.1076/ceyr.23.3.185.5465.
To evaluate differences of phagocytic capacities in rat retinal pigment epithelial (RPE) cells, iris pigment epithelial (IPE) cells, and basic fibroblast growth factor (bFGF) cDNA-transfected IPE (bFGF-IPE) cells in vitro.
The RPE cells and IPE cells were isolated from adult Long Evans rats' eyes. The bFGF cDNA was transfected to the IPE using lipofection method. The bovine photoreceptor outer segments (POS) were isolated and labeled with Alexa 488 dye (fluorescent marker, EX: 494 nm, EM: 519 nm), and were applied to the cultured RPE, IPE, vector-IPE, and bFGF-IPE for 20 hours. The amount of phagocytosed POS in cells was photographed by fluorescence microscopy with fluorescein isothiocyanate (FITC) filter, and expressed as the ratio of the occupying area of fluorescence to the area of pigment epithelial cells in each observing field.
The phagocytic capacity of IPE and vector-IPE were about 70% of RPE, and that of bFGF-IPE increased to 150% of IPE. This increase was inhibited by pretreatment with anti-bFGF antibody to bFGF-IPE.
IPE showed phagocytic capacity, but it reached only 70% of that in RPE. The stable expression of bFGF promoted its function.
评估大鼠视网膜色素上皮（RPE）细胞、虹膜色素上皮（IPE）细胞以及碱性成纤维细胞生长因子（bFGF）cDNA转染的IPE（bFGF-IPE）细胞在体外吞噬能力的差异。
从成年Long Evans大鼠眼中分离RPE细胞和IPE细胞。采用脂质转染法将bFGF cDNA转染至IPE细胞。分离牛光感受器外段（POS）并用Alexa 488染料（荧光标记物，激发波长：494 nm，发射波长：519 nm）进行标记，然后将其应用于培养的RPE、IPE、载体转染的IPE（载体-IPE）和bFGF-IPE细胞20小时。用异硫氰酸荧光素（FITC）滤光片通过荧光显微镜拍摄细胞中吞噬的POS数量，并表示为每个观察视野中荧光占据面积与色素上皮细胞面积的比值。
IPE和载体-IPE的吞噬能力约为RPE的70%，而bFGF-IPE的吞噬能力增加至IPE的150%。用抗bFGF抗体预处理bFGF-IPE可抑制这种增加。
IPE显示出吞噬能力，但仅达到RPE吞噬能力的70%。bFGF的稳定表达促进了其功能。
Zotero 插件