Engelmann K, Bednarz J, Böhnke M
Universitäts-Augenklinik Hamburg.
Ophthalmologe. 1999 Sep;96(9):555-62. doi: 10.1007/s003470050452.
The human corneal endothelium has a limited proliferative capacity in vivo. Until now it has only been possible to replace damaged endothelium by transplantation of a donor cornea. After establishing methods for the isolation and in vitro cultivation of human corneal endothelial cells, transplantation of these cells my be an alternative therapeutic option.
In this review methods for the in vitro cultivation of human corneal endothelial cells and their transplantation on the Descemet membrane of donor corneas are described.
In vitro proliferation of human adult corneal endothelial cells was achieved by the development of defined cell culture conditions, including supplementation of culture medium with specified growth factors and substances. Dependent on the culture conditions, as well as independent of them, in vitro cultured endothelial cells showed phenotypic changes and different proliferative behavior. Thus, molecular biological examinations revealed a different expression pattern of growth factor receptors in fibroblast-like endothelial cells (dedifferentiated) compared to typical endothelial cells (differentiated). Moreover, the proliferative capacity of the cells differed, dependent on their corneal location. Cells isolated from the peripheral part of donor corneas have a higher proliferative capacity than cells obtained from the central part. The propagation of corneal endothelial cells in vitro offered the possibility of their transplantation on donor corneas in an in vitro model. After transplantation, these cells formed a monolayer whose morphology and cell density depended on the differentiation of the cells. DNA synthesis was predominantly detectable in cells of the corneal periphery.
Our findings are the basis of the following hypothesis: the periphery of the cornea represents a regenerative zone of the corneal endothelium. The fact that early after transplantation corneal endothelial cells form a monolayer on the natural extracellular matrix (ECM), which shows contact inhibition, suggests that inhibitory factors are released by the Descemet membrane that influence the proliferation of the cells. Further studies on the regulation of the proliferation and differentiation of human corneal endothelial cells in vitro and after transplantation might offer the possibility to establish a selective procedure for the treatment of corneal endothelial cell loss in the near future.
人角膜内皮细胞在体内的增殖能力有限。到目前为止,只能通过移植供体角膜来替换受损的内皮细胞。在建立了人角膜内皮细胞的分离和体外培养方法后,这些细胞的移植可能成为一种替代治疗选择。
本综述描述了人角膜内皮细胞的体外培养方法及其在供体角膜后弹力层上的移植。
通过确定细胞培养条件,包括在培养基中添加特定生长因子和物质,实现了人成年角膜内皮细胞的体外增殖。取决于培养条件以及与之无关的因素,体外培养的内皮细胞表现出表型变化和不同的增殖行为。因此,分子生物学检查显示,与典型内皮细胞(分化型)相比,成纤维细胞样内皮细胞(去分化型)中生长因子受体的表达模式不同。此外,细胞的增殖能力也有所不同,这取决于它们在角膜中的位置。从供体角膜周边分离的细胞比从中央获得的细胞具有更高的增殖能力。角膜内皮细胞在体外的增殖为其在体外模型中移植到供体角膜上提供了可能性。移植后,这些细胞形成了单层,其形态和细胞密度取决于细胞的分化程度。DNA合成主要在角膜周边的细胞中检测到。
我们的研究结果是以下假设的基础:角膜周边代表角膜内皮的再生区域。移植后早期角膜内皮细胞在天然细胞外基质(ECM)上形成单层并表现出接触抑制这一事实表明,后弹力层释放影响细胞增殖的抑制因子。对人角膜内皮细胞在体外和移植后增殖与分化调控的进一步研究可能在不久的将来为建立治疗角膜内皮细胞丢失的选择性程序提供可能性。