Fan Z J, Wei H R, Wang A
Department of Ophthalmology, Union Hospital, Tongji Medical University, Wuhan, P.R. China.
Graefes Arch Clin Exp Ophthalmol. 1996 Jan;234(1):64-6. doi: 10.1007/BF00186521.
The interaction between different cells plays an important role in many physiological and pathological processes. Since the proliferation of fibroblasts is very much involved in the pathogenesis of eye diseases such as proliferative vitreoretinopathy, the failure of filtration in glaucoma surgery, etc., we attempted to ascertain whether iris pigment epithelial cells (IPE) have some modulating effect on fibroblast proliferation.
Human IPE were explanted and the third-passage culture was transferred into serum-free RPMI-1640 medium. After 48 h of further incubation, the medium was collected and submitted to centrifugation; the supernatant was used as the conditioned medium of IPE (IPE-CM). Cultured fibroblasts from Tenon's capsule were seeded in a 96-well plate and incubated with IPE-CM in different concentrations. The proliferation of fibroblasts was estimated by thymidine incorporation and cell counting.
The incorporation of tritiated thymidine by fibroblasts was reduced to 56.68% of baseline with 1:16 diluted IPE-CM and to 13.63% and 8.20%, respectively, with 1:8 and 1:2 diluted IPE-CM. These findings were in good accordance with the results of cell counting, performed in parallel. SDS-PAGE of IPE-CM revealed two specific bands with molecular weight 65 kDa and 40 kDa.
IPE-CM showed an obvious dose-dependent inhibitory effect on fibroblast proliferation and was presumed to contain some active factors contributing to this effect.
不同细胞之间的相互作用在许多生理和病理过程中起着重要作用。由于成纤维细胞的增殖在增殖性玻璃体视网膜病变、青光眼手术滤过失败等眼部疾病的发病机制中密切相关，我们试图确定虹膜色素上皮细胞（IPE）是否对成纤维细胞增殖具有某种调节作用。
将人IPE进行外植培养，第三代培养物转移至无血清的RPMI - 1640培养基中。进一步孵育48小时后，收集培养基并进行离心；上清液用作IPE的条件培养基（IPE - CM）。将来自Tenon囊的培养成纤维细胞接种于96孔板中，并用不同浓度的IPE - CM进行孵育。通过胸腺嘧啶核苷掺入法和细胞计数评估成纤维细胞的增殖。
用1:16稀释的IPE - CM时，成纤维细胞对氚标记胸腺嘧啶核苷的掺入量降至基线的56.68%，用1:8和1:2稀释的IPE - CM时，分别降至13.63%和8.20%。这些结果与同时进行的细胞计数结果高度一致。IPE - CM的SDS - PAGE显示出两条分子量分别为65 kDa和40 kDa的特异性条带。
IPE - CM对成纤维细胞增殖显示出明显的剂量依赖性抑制作用，并推测其含有一些导致该作用的活性因子。
