Academic Unit of Ophthalmology, University of Bristol, Bristol Eye Hospital, Bristol, UK.
Ophthalmic Res. 2010;43(1):33-42. doi: 10.1159/000246576. Epub 2009 Oct 14.
The traditional medium for storing corneas in European Eye Banks is Gibco's MEM (Eagle's) with Earle's salts and Hepes containing 2% fetal calf serum, glutamine and antimycotics. Although serum-free MegaCell MEM has been reported to be more suitable for this purpose, it contains components of animal origin that potentially pose health risks to corneal recipients. The possibility of removing or replacing these components has therefore been investigated.
A MegaCell basal medium (DME) and a formulation of this (MegaCell DCS), which contains no components of animal origin, have been prepared. The viability of the endothelial, epithelial and stromal cells of corneas held in these media has been assessed, their stress levels monitored and water content determined.
The endothelial cell count and morphology of corneas held in MegaCell DME and DCS for 30 days remained little changed. Their epithelial and stromal cells also retained their ability to proliferate in culture. Neither DME nor DCS prevented corneal swelling but the lack of endogenous MMP-2 activation indicated that the corneas were not subject to metabolic stress.
MegaCell DCS is an animal product-free medium formulated for corneal storage. The quality of corneas held in this medium is similar to those held in MegaCell MEM.
在欧洲眼库中,储存角膜的传统介质是含有 2%胎牛血清、谷氨酰胺和抗真菌药物的 Gibco 的 MEM(Eagle's)和 Earle's 盐以及 Hepes。尽管无血清的 MegaCell MEM 已被报道更适合用于此目的,但它含有潜在对角膜接受者构成健康风险的动物源性成分。因此,已经研究了去除或替换这些成分的可能性。
已经制备了 MegaCell 基础培养基(DME)和该培养基的一种配方(MegaCell DCS),其中不含任何动物源性成分。评估了这些介质中保存的角膜内皮细胞、上皮细胞和基质细胞的活力,监测了它们的应激水平并确定了水含量。
在 MegaCell DME 和 DCS 中保存 30 天的角膜内皮细胞计数和形态变化不大。它们的上皮细胞和基质细胞也保留了在培养中增殖的能力。DME 和 DCS 都不能防止角膜肿胀,但缺乏内源性 MMP-2 激活表明角膜没有受到代谢应激。
MegaCell DCS 是一种不含动物产品的培养基,专为角膜储存而配制。在这种培养基中保存的角膜质量与在 MegaCell MEM 中保存的角膜相似。