Effect of human autologous serum and fetal bovine serum on human corneal epithelial cell viability, migration and proliferation in vitro

被引:21
作者
Wu, Ming-Feng [1 ]
Stachon, Tanja [1 ]
Seitz, Berthold [1 ]
Langenbucher, Achim [2 ]
Szentmary, Nora [1 ,3 ]
机构
[1] Saarland Univ, Med Ctr, Dept Ophthalmol, D-66424 Homburg, Germany
[2] Saarland Univ, Expt Ophthalmol, D-66424 Homburg, Germany
[3] Semmelweis Univ, Dept Ophthalmol, H-1085 Budapest, Hungary
关键词
autologous serum; eye drops; serum concentration; migration; proliferation; viability; human corneal epithelial cells; FIBROBLAST-GROWTH-FACTOR; OCULAR SURFACE DISEASE; DRY EYE SYNDROME; ARTIFICIAL TEARS; DROPS; CULTURE; KERATOCONJUNCTIVITIS; EQUIVALENTS; CROSSOVER; EYEDROPS;
D O I
10.18240/ijo.2017.06.12
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
AIM: To analyze the concentration-dependent effects of autologous serum (AS) and fetal bovine serum (FBS) on human corneal epithelial cell (HCEC) viability, migration and proliferation.. METHODS: AS was prepared from 13 patients with non-healing epithelial defects Dulbecco's modified eagle medium/Ham's F12 (DMEM/F12) with 5% FBS, 0.5% dimethyl sulphoxide (DMSO), 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media: DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTT, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay (ELISA) BrdU kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines. RESULTS: HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse (P=0.001, P=0.023) compared to baseline and significantly better at 15% FBS (P=0.003) concentrations. HCEC migration was significantly worse (P <= 0.007) and HCEC proliferation significantly better (P<0.001) in all concentration groups compared to baseline. CONCLUSION: For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.
引用
收藏
页码:908 / 913
页数:6
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