Isolation efficiency of collagenase and EDTA for the culture of corneal endothelial cells.

IF 1.4 3区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Vision Pub Date : 2022-10-02 eCollection Date: 2022-01-01

Kim Santerre, Stéphanie Proulx

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引用次数: 0

Abstract

Purpose: Tissue engineering of the corneal endothelium, as well as cell therapy, has been proposed as an alternative approach for the treatment of corneal endotheliopathies. These approaches require in vitro amplification of functional corneal endothelial cells (CECs). The goal of this study was to compare two common isolation methods, collagenase A and EDTA (EDTA), and determine whether they influence cell viability, morphology, and barrier function.

Methods: Human eye bank research-grade corneas were used to isolate and cultivate CECs. All donors were more than 40 years old. Two Descemet membranes from the same donor were used separately to compare the collagenase A and EDTA cell isolation methods. The number of isolated cells, cell viability, morphology, and barrier functionality were compared.

Results: A higher isolation efficiency of viable CECs and a higher circularity index (endothelial morphology) were obtained using collagenase A. Passage 3 cells presented similar barrier functionalities regardless of the isolation method.

Conclusions: This study showed that isolation of CECs using collagenase A yields higher isolation efficiency than EDTA, delaying the loss of endothelial morphology for early passage cells.

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胶原酶和EDTA在角膜内皮细胞培养中的分离效果。

目的:角膜内皮组织工程和细胞治疗已被提出作为治疗角膜内皮病变的另一种方法。这些方法需要功能性角膜内皮细胞(CECs)的体外扩增。本研究的目的是比较两种常见的分离方法，胶原酶A和EDTA (EDTA)，并确定它们是否影响细胞活力、形态和屏障功能。方法:采用人眼库研究级角膜分离培养CECs。所有的捐赠者都在40岁以上。分别使用同一供体的两种Descemet膜，比较胶原酶A和EDTA细胞的分离方法。比较分离细胞的数量、细胞活力、形态和屏障功能。结果:采用胶原酶A可获得较高的活性CECs分离效率和较高的圆形指数(内皮细胞形态)，无论采用何种分离方法，传代3细胞均具有相似的屏障功能。结论:本研究表明，使用胶原酶A分离CECs比使用EDTA分离效率更高，可以延缓早期传代细胞内皮形态的丧失。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Vision 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Molecular Vision is a peer-reviewed journal dedicated to the dissemination of research results in molecular biology, cell biology, and the genetics of the visual system (ocular and cortical). Molecular Vision publishes articles presenting original research that has not previously been published and comprehensive articles reviewing the current status of a particular field or topic. Submissions to Molecular Vision are subjected to rigorous peer review. Molecular Vision does NOT publish preprints. For authors, Molecular Vision provides a rapid means of communicating important results. Access to Molecular Vision is free and unrestricted, allowing the widest possible audience for your article. Digital publishing allows you to use color images freely (and without fees). Additionally, you may publish animations, sounds, or other supplementary information that clarifies or supports your article. Each of the authors of an article may also list an electronic mail address (which will be updated upon request) to give interested readers easy access to authors.