期刊
BIOMATERIALS
卷 28, 期 27, 页码 3928-3935出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2007.05.009
关键词
cell adhesion; cell culture; cell spreading; endothelial cell; integrin; shear
资金
- NHLBI NIH HHS [R01 HL044972-17, HL-44972, R01 HL044972] Funding Source: Medline
- NIGMS NIH HHS [GM8555, T32 GM008555-13, T32 GM008555] Funding Source: Medline
A necessary condition for endothelialization of small diameter grafts is rapid and firm adhesion of endothelial cells upon exposure to flow. To retain integrins on the cell surface, we assessed the effects of trypsin concentration, the duration of trypsin incubation, and trypsin neutralization methods on endothelial cell adhesion. Human umbilical vein endothelial cells which were detached using 0.025% trypsin for 5 min and seeded onto glass pretreated with fibronectin had close to 100% cell retention when shear stresses as high as 200 dyn/cm(2) were applied for 2 min. An equivalent level of cell retention was observed on fibronectin coated Teflon-AF(TM) for shear stresses up to 60dyn/cm(2) applied for 4h. Using 0.025% trypsin, initial cell spreading and cell surface alpha(5)beta(1) integrins were increased relative to cells treated with 0.5% trypsin. After 1 h of attachment, focal adhesions formed when low trypsin concentrations were used but were less evident with high trypsin concentrations. These results showed that low trypsin concentrations produced faster spreading, a higher number of intact integrins, and rapid focal adhesion formation. (C) 2007 Elsevier Ltd. All rights reserved.
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