Centre de recherche du Centre hospitalier universitaire (CHU) de Québec - Université Laval, axe médecine régénératrice, Hôpital du Saint-Sacrement, Québec, QC, Canada.
Département d'Ophtalmologie et d'oto-rhino-laryngologie-chirurgie cervico-faciale, Faculté de médecine, Université Laval, Québec, QC, Canada.
Mol Vis. 2022 Oct 2;28:331-339. eCollection 2022.
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.
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.
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.
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.
角膜内皮组织工程以及细胞疗法已被提议作为治疗角膜内皮病变的一种替代方法。这些方法需要体外扩增功能性角膜内皮细胞（CEC）。本研究的目的是比较两种常见的分离方法，即胶原酶 A 和 EDTA（EDTA），并确定它们是否会影响细胞活力、形态和屏障功能。
使用人眼库研究级角膜分离和培养 CEC。所有供体年龄均超过 40 岁。从同一位供体中使用两块 Descemet 膜分别比较胶原酶 A 和 EDTA 细胞分离方法。比较分离细胞的数量、细胞活力、形态和屏障功能。
使用胶原酶 A 可获得更高的活 CEC 分离效率和更高的圆形指数（内皮形态）。第 3 代细胞无论采用哪种分离方法，其屏障功能均相似。
本研究表明，使用胶原酶 A 分离 CEC 可获得比 EDTA 更高的分离效率，延缓早期传代细胞内皮形态的丧失。
