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首页> 外文期刊>Biotechnology Letters >A novel human donor cornea preservation cocktail incorporating a thermo-reversible gelation polymer (TGP), enhancing the corneal endothelial cell density maintenance and explant culture of corneal limbal cells
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A novel human donor cornea preservation cocktail incorporating a thermo-reversible gelation polymer (TGP), enhancing the corneal endothelial cell density maintenance and explant culture of corneal limbal cells

机译：一种新型人体供体角膜保存鸡尾酒，其掺入热可逆凝胶化聚合物（TGP），增强角膜内皮细胞密度维持和角膜内皮细胞的脱钙培养物

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Purpose McCarey-Kaufman's (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media. Methods Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 degrees C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days. Results MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization. Conclusion TGP reconstituted with MK and Optisol-GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.
机译：目的，McCarey-Kaufman（MK）中和Optisol-GS媒体是人类供体角膜保存最常用的媒体。在该研究中，我们评估了使用热可逆凝胶化聚合物（TGP）在这两个培养基中的丢弃的人供体角膜的保存疗效。方法使用从死者供体中收集的十三个人角膜按钮，其由于低内皮细胞密度（ECD）而被丢弃。它们以四组：MK培养基，MK培养基，具有TGP，TGP的Optisol-GS，在4℃下为96小时。切片灯检查和镜面显微镜检查。然后使用来自这些角膜的角膜斜腹组织使用外蛋白方法，另一种没有TGP支架，21天。结果MK + TGP和OPTISOL-GS + TGP保存的玉米羚比没有TGP更好，其通过维持ECD，其在Optisol-GS + TGP中显着高于Mk + TGP（P值= 0.000478）和剩余的角膜厚度96小时相同。可以从仅在MK + TGP和Optisol-GS + TGP中储存的玉米体生长活角膜上皮细胞。在培养过程中，通过RT-PCR表征确认了TGP支架帮助维持天然上皮表型和祖细胞/祖细胞生长。结论TGP重构MK和OPTISOL-GS培养基，在相对较高的ECD维持和角膜斜坡组织的体外培养结果方面，对人角膜按钮的保存更好。该方法有可能成为标准供体角膜运输保存方法，并且在进一步验证时也可以扩展到其他组织或器官运输。

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