Effects of growth factors (EGF, PDGF-BB and TGF-β1) on cultured equine epithelial cells and keratocytes:: implications for wound healing
Effects of growth factors (EGF, PDGF-BB and TGF-β1) on cultured equine epithelial cells and keratocytes:: implications for wound healing
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DOI：
10.1046/j.1463-5224.2003.00296.x
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发表时间：
2003-09-01
影响因子：
1.6
通讯作者：
Provost, PJ
中科院分区：
文献类型：
--
作者：
Haber, M;Cao, ZY;Provost, PJ
目的角膜创伤愈合过程中，生长因子（包括PDGF-BB、EGF和TGF-β 1）作为成纤维细胞和上皮细胞的有效介质的生理机制尚不清楚。本研究的目的是确定马上皮细胞和角膜上皮细胞的培养方法，并探讨外源性生长因子如何影响这两种细胞类型的增殖。程序细胞培养建立从健康的角膜收获后立即安乐死的马，并保持使用标准的组织培养方案。为了确定PDGF-BB、EGF、TGF-β 1、角化细胞（1 × 10（5）/孔）和上皮细胞（2 × 10（5）/孔）各自在12孔板中培养，并分别暴露于生长因子。将细胞暴露于浓度为0至50 ng/mL的EGF、0至75 ng/mL的PDGF-BB和0至10 ng/mL的TGF-β 1。3 H-胸腺嘧啶核苷法检测细胞增殖，方差分析和Tukey 'sHSD检验检测细胞生长差异（P < 0.05）。EGF分别在2.5ng/mL和5 ng/mL时对角膜细胞和上皮细胞的刺激作用最大。PDGF-BB分别在50 ng/ mL和5 ng/mL时最大程度地刺激角膜细胞和上皮细胞。TGF-β1在5 ng/mL和10 ng/mL时对角膜基质细胞有抑制作用，在1 ng/mL和2ng/mL时对上皮细胞有抑制作用。PDGF-BB和EGF刺激上皮细胞和角膜基质细胞的增殖，而TGF-β1抑制上皮细胞和角膜基质细胞的增殖。这些生长因子可能在马角膜的维护和修复中发挥作用。
Objective The physiologic mechanisms involving growth factors, including PDGF-BB, EGF, and TGF-beta1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types.Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF-BB, EGF, TGF-beta1, keratocyteS (1 X 10(5)/well) and epithelial cells (2 X 10(5) /well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 5 0 ng/mL; PDGF-BB between 0 and 7 5 ng/mL; and TGF-beta1 between 0 and 10 ng/mL. Cell proliferation was measured using 3 H-thymidine assay and differences in growth determined using ANOVA and Tukey's HSD test (P < 0.05).Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 2 5 ng/mL and 5 ng/mL, respectively. PDGF-BB maximally stimulated keratocytes and epithelial cells at 50 ng/ mL and 5 ng/mL, respectively. TGF-β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL.Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF-BB and EGF stimulate, while TGF-β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.