Ellis D G, Cheng Q, Lee D A
Jules Stein Eye Institute, Department of Opththalmology, UCLA School of Medicine 90095-7004, USA.
Curr Eye Res. 1996 Jan;15(1):27-35. doi: 10.3109/02713689609017608.
This study was performed to develop and improve a completely defined in vitro ocular wound-healing model of fibroblast proliferation for glaucoma filtration surgery. This model is essential for the investigation of protein-sensitive drugs and cytokines. Tenon's capsule fibroblasts in their third passage were incubated overnight, washed free of serum, and fed defined media, Aim V or Clonetics FBM serum-free medium containing platelet-derived growth factor, basic fibroblast growth factor, epidermal growth factor, or fibronectin at various dilutions and in combinations at optimum concentrations. Proliferation was measured by 3H-thymidine incorporation at 1, 3, and 7 days. Morphology was compared to controls fed Minimum Essential Medium + 10% serum. Single factors stimulated the greatest amount of thymidine uptake on day 3. Optimum concentrations were epidermal growth factor at 5 ng/ml, basic fibroblast growth factor at 10 ng/ml and platelet-derived growth factor at 20 ng/ml. Identical combinations of factors stimulated nearly twice the thymidine uptake in Clonetics medium as in Aim V. Epidermal growth factor activity was inhibited by either basic fibroblast growth factor or platelet-derived growth factor. Basic fibroblast growth factor and platelet-derived growth factor together produced a less than additive effect. The performance of either serum-free medium may be improved by the addition of basic fibroblast growth factor or platelet-derived growth factor. The optimum serum-free medium (Clonetics FBM) with growth factors was unable to stimulate proliferation as much as Minimum Essential Medium + 10% NBS, but was successful in maintaining viability during the 7 day test period.
本研究旨在开发并改进一种完全确定的体外成纤维细胞增殖性青光眼滤过手术眼部伤口愈合模型。该模型对于研究蛋白敏感性药物和细胞因子至关重要。将第三代的眼球筋膜成纤维细胞进行过夜培养，洗去血清，然后用确定的培养基、Aim V或Clonetics FBM无血清培养基喂养，这些培养基含有不同稀释度及最佳浓度组合的血小板衍生生长因子、碱性成纤维细胞生长因子、表皮生长因子或纤连蛋白。在第1、3和7天通过3H-胸苷掺入法测量增殖情况。将形态与用最低必需培养基+10%血清喂养的对照进行比较。单一因子在第3天刺激的胸苷摄取量最大。最佳浓度为5 ng/ml的表皮生长因子、10 ng/ml的碱性成纤维细胞生长因子和20 ng/ml的血小板衍生生长因子。相同的因子组合在Clonetics培养基中刺激的胸苷摄取量几乎是Aim V中的两倍。碱性成纤维细胞生长因子或血小板衍生生长因子可抑制表皮生长因子的活性。碱性成纤维细胞生长因子和血小板衍生生长因子共同产生的效果小于相加效应。添加碱性成纤维细胞生长因子或血小板衍生生长因子可改善任何一种无血清培养基的性能。含有生长因子的最佳无血清培养基（Clonetics FBM）刺激增殖的能力不如最低必需培养基+10%新生牛血清，但在7天测试期内成功维持了细胞活力。
