Establishment of optirmal culture media in corneal epithelial wound healing models

Q3 Biochemistry, Genetics and Molecular Biology Journal of Cellular Biotechnology Pub Date : 2021-12-20 DOI:10.3233/jcb-210039
D. McCanna
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Abstract

BACKGROUND: Wound healing needs to occur after injury to prevent vision loss. Models of wound healing need to be optimized to assure treatments for corneal wounds can be developed in vitro prior to investigating with in vivo studies. OBJECTIVE: The purpose of this study was to establish the optimum media to use as a control solution in wound healing models. METHODS: Immortalized human corneal epithelial cells were cultured in different growth media using a scratch and exclusion zone model. The effect of normoxic and hypoxic conditions on tight junctional integrity and metabolic activity of cells grown in different growth medium were also investigated. RESULTS: Wound healing with DMEMF12 media was significantly faster than both Keratinocyte serum-free media (p <  0.05) and EpiLife (p <  0.05) after 10 hours recovery under normoxic or hypoxic conditions using the scratch model and 9 days after wounding using the exclusion zone technique (p <  0.05). Using the culture media DMEMF12, cells stained for abundant ZO-1, Cx43 and had a high metabolic activity indicating significant epithelial barrier formation, gap junction formation and high cell viability. CONCLUSIONS: DMEMF12 led to superior wound healing under hypoxic and normoxic conditions and in two different wound healing models.
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角膜上皮伤口愈合模型光热培养基的建立
背景:受伤后需要进行伤口愈合,以防止视力下降。需要优化伤口愈合模型,以确保在进行体内研究之前,可以在体外开发角膜伤口的治疗方法。目的:本研究的目的是建立在伤口愈合模型中用作对照溶液的最佳培养基。方法:采用划痕和禁区模型,在不同的生长培养基中培养永生人角膜上皮细胞。还研究了常氧和缺氧条件对不同生长培养基中生长的细胞紧密连接完整性和代谢活性的影响。结果:DMEMF12培养基的伤口愈合速度明显快于无角质形成细胞培养基(p <  0.05)和EpiLife(p <  0.05)在使用划痕模型的常氧或缺氧条件下恢复10小时后和使用禁区技术的创伤后9天(p <  0.05)。使用培养基DMEMF12,细胞对丰富的ZO-1、Cx43进行染色,并具有高代谢活性,表明显著的上皮屏障形成、间隙连接形成和高细胞活力。结论:DMEMF12在低氧和常氧条件下以及在两种不同的伤口愈合模型中都能导致良好的伤口愈合。
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来源期刊
Journal of Cellular Biotechnology
Journal of Cellular Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
0.70
自引率
0.00%
发文量
13
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