评估 Megacell MEM 作为用于移植目的角膜的储存介质。

Evaluation of Megacell MEM as a storage medium for corneas destined for transplantation.

机构信息

University of Bristol, Academic Unit of Ophthalmology, Bristol Eye Hospital, Bristol, UK.

出版信息

Ophthalmic Res. 2010;43(1):18-25. doi: 10.1159/000246574. Epub 2009 Oct 14.

DOI:10.1159/000246574

PMID:19829008

Abstract

PURPOSE

Gibco's Minimum Essential Medium with Earle's salts and HEPES supplemented with glutamine, antibiotics (EB MEM) and 2% foetal calf serum (FCS) is used in European eye banks to store corneas. Although FCS is important to endothelial cell survival in this medium, it is a potential biohazard. Megacell MEM, formulated to reduce the FCS requirement of cells by a factor of 5, has therefore been evaluated as a corneal storage medium.

METHODS

Corneal stromal and epithelial cells were incubated in Megacell MEM (serum-free or 2% FCS) to assess their viability in these media. Endothelial cell densities of paired corneas held in either EB MEM 2% FCS or Megacell MEM (serum-free or 2% FCS) were measured over 5 weeks. Discs subsequently punched from the centre of these corneas were weighed, dried and reweighed to determine hydration levels.

RESULTS

Both corneal stromal and epithelial cells proliferated in Megacell MEM 2% FCS. Relative to EB MEM, 2% FCS Megacell MEM prolonged the viability of corneal endothelial cells and improved their morphological appearance, irrespective of whether it contained FCS or not. This was independent of corneal swelling.

CONCLUSION

Serum-free Megacell MEM is a better storage medium than EB MEM 2% FCS for corneas destined for transplantation.

摘要

目的

吉布斯最小必需培养基（含 Earle 盐和 HEPES），添加谷氨酰胺、抗生素（EB MEM）和 2%胎牛血清（FCS），用于欧洲眼库储存角膜。虽然 FCS 对该培养基中的内皮细胞存活很重要，但它是一种潜在的生物危害。Megacell MEM 通过将细胞对 FCS 的需求降低 5 倍来进行配方，因此被评估为一种角膜储存介质。

方法

将角膜基质和上皮细胞在 Megacell MEM（无血清或 2% FCS）中孵育，以评估它们在这些培养基中的生存能力。在 EB MEM 2% FCS 或 Megacell MEM（无血清或 2% FCS）中保存的配对角膜的内皮细胞密度在 5 周内进行测量。随后从这些角膜的中心打孔，称重、干燥并再次称重以确定水合水平。