The Use of Mild Trypsinization Conditions in the Detachment of Endothelial Cells to Promote Subsequent Endothelialization on Synthetic Surfaces.
From: Department of Biomedical Engineering, Duke University, 136 Hudson Hall, Campus Box 90281, Durham, NC 27708-0281, USA.
Biomaterials
- Publish Date: Sep 2007
- ISSN: 0142-9612
- Volume: 28
- Issue: 27
- Pages: 3928-35
- Medium: Print
- Language: English
- Citation (JAMA): Brown Melissa A, Wallace Charles S, Anamelechi Charles C, et al. The Use of Mild Trypsinization Conditions in the Detachment of Endothelial Cells to Promote Subsequent Endothelialization on Synthetic Surfaces.. Biomaterials Sep 2007;28:3928-35
Abstract
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/cm2 were applied for 2 min. An equivalent level of cell retention was observed on fibronectin coated Teflon-AF for shear stresses up to 60 dyn/cm2 applied for 4h. Using 0.025% trypsin, initial cell spreading and cell surface alpha5beta1 integrins were increased relative to cells treated with 0.5% trypsin. After 1h 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.
Mesh Headings (Keywords): Cell Adhesion, Cell Culture Techniques, Cell Proliferation, Cells, Cultured, Endothelial Cells, Endothelium, Vascular, Humans, Trypsin
Check for Full Text / PubMed Unique Identifier (PMID): 17570483
This abstract is part of PubMed, a service of the U.S. National Library of Medicine. PubMed includes more than 17 million citations from MEDLINE and other life science journals for biomedical articles. See Copyright and Disclaimers.
Linked medical terms appearing on this page are added by Healia to help readers find more information and are not part of the original PubMed document.
The data herein was last updated on July 8th, 2008 and may not reflect the most current and accurate data available from NLM.