The Effects of Epidermal Growth Factor and Chondroitin Sulfate on the Animal Corneal Endothelial Cell Culture

表皮生長因子和硫酸軟骨素對動物角膜內皮細胞培養的影響

李弘志(Hong-Ju Lee) ； 林昌平(Chang-Ping Lin) ； 陳振武(Chen-Wu Chen)

《The Kaohsiung Journal of Medical Sciences》 7卷12期 (1991/12) Pp. 614-621

https://doi.org/10.6452/KJMS.199112.0614

摘要

本文以豬的角膜內板細胞行細胞培養，研究不同濃度的表皮生長因子（EGF）及硫酸軟骨素（CDS）對培養細胞生長的影響。實驗共分七組，在Earle’s基本培養液中加上各種濃度的添加物,分別是：〔1#〕為僅加入EMEM；〔2#〕為EMEM＋EGF（10ng/ml）；〔3#〕為EMEM+EGF（100ng/ml）；〔4#〕為EMEM+CDS（1mg/ml）；〔5#〕為EMEM+CDS（25mg/ml）；〔6#〕為EMEM+EGF（10ng/ml）+CDS（1mg/ml）及〔7#〕為EMEM+EGF（100ng/ml）+CDS（25mg/ml）。結果顯示各組的培殖時間分別是：〔1#〕為47.66小時；〔2#〕為50.44小時；〔3#〕為39.05小時；〔4#〕為47.66小時；〔5#〕為75.25小時；〔6#〕為52.96小時；及〔7#〕59.97小時。而達融合（confluence）的時間分別是〔1#〕為264小時；〔2#〕為264小時；〔3#〕為216小時；〔4#〕為264小時；〔5#〕為384小時；
〔6#〕為288小時；及〔7#〕為312小時。第3#組的生長速度明顯地比其他各組為快，而第5#組的細胞生長有明顯遲滯作用，此表示高濃度的表皮生長因子
（100ng/ml）對豬角膜內板細胞培養有促進作用，而高濃度的硫酸軟骨素（25mg/ml）則對內板細胞培養的生長有遲滯作用。低濃度的表皮生長因子
（100ng/ml）及低濃度的硫酸軟骨素（1mg/ml）對豬角膜內皮細胞培養的影響並不顯著。在細胞生長過程中，第3#組的分裂能力有明顯加強，可見相當多的雙核及多核仁細胞，並可見許多核周圍顆粒，而第7#的細胞外形較特殊，其細胞呈現較扁平的外形，且仪貼於培養皿的底部。角膜內皮細胞培養乃是細胞移植及角膜內皮細胞加強作用的基礎，這方面的研究，將來或許能解決捐贈眼球來源不足的問題。

並列摘要

In order to investigate the effects of different culture media containing various concentrations of epidermal growth factor (EGF) and chondroitin sulfate (CDS) on the growth of cultured corneal endothelial cells were used for this study. The cells were divided into 7 groups and each group was cultured with a medium containing: 1. Eagle’s minimal essential medium with Earle’s salt (EMEM) only; 2. EMEM + EGF (10 ng/ml); 3. EMEM + EGF (100 ng/ml); 4. EMEM + CDS (1 mg/ ml); 5. EMEM + CDS (25 mg/ml); 6. EMEM + EGF (10 ng/ml) + CDS (1 mg/ml); 7. EMEM + EGF (100ng/ml) + CDS (25mg/ml),respectively.
The results are shown below:
(1)High concentration of EGF (100 ng/ml) stimulated the growth of pig corneal endothelial cells, shortened doubling time and the cells reached confluence earliest in group 3. But a low concentration of EGF (10 ng/ml) showed no effects on the growth of pig corneal endothelial cells.
(2)A high concentration of CDS (25 mg/ml) might retard the growth of pig corneal endothelial cell, but a low concentration of CDS (1 mg/ml) has no retarding effects on the growth of pig corneal endothelial cells.
(3)A high concentration of EGF could “antagonize” the CDS induced growth retarding effects of endothelial cells.
(4)Those cells cultured with a medium containing high concentrations of EGF had more mitotic activity, including many prominent binuclear and polynucleolar cells.
(5)The cells cultured with a medium containing high concentrations of CDS showed a more flat-shaped morphology under observation with a phase contrast microscope.
The results of this study may help to improve the technique for corneal preservation.

並列關鍵字

epidermal growth factor ； chondroitin sulfate ； corneal endothelium