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The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces.

Publication ,  Journal Article
Brown, MA; Wallace, CS; Anamelechi, CC; Clermont, E; Reichert, WM; Truskey, GA
Published in: Biomaterials
September 2007

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.

Duke Scholars

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

September 2007

Volume

28

Issue

27

Start / End Page

3928 / 3935

Related Subject Headings

  • Trypsin
  • Humans
  • Endothelium, Vascular
  • Endothelial Cells
  • Cells, Cultured
  • Cell Proliferation
  • Cell Culture Techniques
  • Cell Adhesion
  • Biomedical Engineering
 

Citation

APA
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ICMJE
MLA
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Brown, M. A., Wallace, C. S., Anamelechi, C. C., Clermont, E., Reichert, W. M., & Truskey, G. A. (2007). The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces. Biomaterials, 28(27), 3928–3935. https://doi.org/10.1016/j.biomaterials.2007.05.009
Brown, Melissa A., Charles S. Wallace, Charles C. Anamelechi, Edward Clermont, William M. Reichert, and George A. Truskey. “The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces.Biomaterials 28, no. 27 (September 2007): 3928–35. https://doi.org/10.1016/j.biomaterials.2007.05.009.
Brown MA, Wallace CS, Anamelechi CC, Clermont E, Reichert WM, Truskey GA. The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces. Biomaterials. 2007 Sep;28(27):3928–35.
Brown, Melissa A., et al. “The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces.Biomaterials, vol. 28, no. 27, Sept. 2007, pp. 3928–35. Epmc, doi:10.1016/j.biomaterials.2007.05.009.
Brown MA, Wallace CS, Anamelechi CC, Clermont E, Reichert WM, Truskey GA. The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces. Biomaterials. 2007 Sep;28(27):3928–3935.
Journal cover image

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

September 2007

Volume

28

Issue

27

Start / End Page

3928 / 3935

Related Subject Headings

  • Trypsin
  • Humans
  • Endothelium, Vascular
  • Endothelial Cells
  • Cells, Cultured
  • Cell Proliferation
  • Cell Culture Techniques
  • Cell Adhesion
  • Biomedical Engineering