Assessment of Attachment Factors for Primary Cultured Human Corneal Endothelial Cells
Assessment of Attachment Factors for Primary Cultured Human Corneal Endothelial Cells
复制标题
DOI：
10.1097/ico.0b013e3181a165a3
复制
发表时间：
2009-10-01
期刊：
影响因子：
2.8
通讯作者：
Jun, Albert S.
中科院分区：
文献类型：
--
作者：
Engler, Christoph;Kelliher, Clare;Jun, Albert S.
目的：探讨各种贴壁因子促进原代培养的人角膜内皮细胞贴壁的能力。材料与方法：原代培养的人角膜内皮细胞在24孔板中孵育2小时。Wells已经被商业上可获得的细胞附着改善介质(FNC涂层混合物)、人胶原1、人纤维连接蛋白、纤维连接蛋白/胶原1或聚D-赖氨酸预涂覆。结果：在未预包被贴壁因子的孔洞中孵育2小时后，出现贴壁迹象的细胞百分率为41+/-16%(平均+/-SD)。FNC涂层混合物、1号胶原蛋白和纤维连接蛋白/I型胶原蛋白在2小时后明显增加细胞贴壁的形态特征。细胞总损失率以聚D-赖氨酸组最高，未加贴壁因子处理组最高。在不使用任何贴壁因子的情况下，漂洗后67+/-19%的细胞仍然存在。结论：I型胶原、I型胶原/纤维连接蛋白、FNC涂层混合物可显著促进人角膜内皮细胞在组织培养板上的铺展。FNC涂层混合物显著减少了因漂洗而造成的细胞损失。在不使用任何附着因子的情况下，67%的细胞在漂洗后仍在原位。
Purpose: To assess the ability of various attachment factors to promote attachment of primary cultured human corneal endothelial cells.Materials and Methods: Primary cultured human corneal endothelial cells (HCEC) were incubated for 2 hours in 24-well plates. Wells had been precoated with commercially available cell attachment improvement media (FNC coating mix), human collagen 1, human fibronectin, fibronectin/collagen 1, or poly-D-lysine. Ratios of cell count before and after rinsing with culture medium and ratios of cells showing morphological signs of spreading to total cells were calculated to measure effectiveness of attachment factors.Results: Incubation of HCEC for 2 hours in wells without pre-coating of attachment factors led to a rate of 41 +/- 16% (mean +/- SD) of cells showing signs of spreading. FNC coating Mix, collagen 1, and fibronectin/collagen I increased significantly the percentage of cells showing morphological features of attachment at 2 hours. Total cell loss was highest with poly-d-lysine and no pretreatment with attachment factor. Without the use of any attachment factor, 67 +/- 19% of cells remained after rinsing. The lowest cell loss was observed with FNC coating mix where 108 +/- 5% of cells remained after rinsing.Conclusion: Collagen I, collagen I/fibronectin, and FNC coating mix significantly enhance the spreading of human corneal endothelial cells to tissue culture plates after 2 hours. FNC coating mix significantly reduces cell loss due to rinsing. Without the use of any attachment factor, 67% of the cells remained in situ after rinsing.