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Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin.

Publication ,  Journal Article
Anamelechi, CC; Clermont, EC; Novak, MT; Reichert, WM
Published in: Langmuir : the ACS journal of surfaces and colloids
May 2009

Surfaces decorated with high affinity ligands can be used to facilitate rapid attachment of endothelial cells; however, standard equilibrium cell detachment studies are poorly suited for assessing these initial adhesion events. Here, a dynamic seeding and cell retention method was used to examine the initial attachment of perfusing human umbilical vein endothelial cells (HUVECs) to bare Teflon-AF substrates, substrates pre-adsorbed with fibronectin alone, or substrates co-pre-adsorbed with two dual-function cell-adhesion ligands: biotinylated fibronectin (bFN) and RGD-streptavidin mutant (RGD-SA). Cell attachment was evaluated as a function of cell trypsinization (integrin digestion), surface protein formulation, and cell perfusion rate. Surfaces co-pre-adsorbed with bFN and RGD-SA showed the highest density of attached cells after 8 min of perfusion and the highest percent retention when subjected to shear flow at 60 dynes/cm2 for 2 min. Surfaces with no ligand treatment showed the lowest cell attachment and retention under flow in all cases. HUVECs trypsinized with mild 0.025% trypsin/ethylenediaminetetraacetic acid (EDTA) showed greater cell adhesion after perfusion and higher percent retention after shear flow than those trypsinized using harsher 0.05% trypsin/EDTA. The preferential affinities of the two dual-function ligands for alpha5beta1 and alphavbeta3 integrins were also examined by surface plasmon resonance (SPR) spectroscopy. The dynamic cell seeding studies confirmed that the dual-function ligand system promotes HUVEC adhesion and retention at short time points when tested using a perfusion assay. SPR studies showed that the two ligands exhibited equal affinity for both alpha5beta1 and alphavbeta3 integrins but that the combined ligands bound more total integrins than the two ligands tested separately.

Duke Scholars

Published In

Langmuir : the ACS journal of surfaces and colloids

DOI

EISSN

1520-5827

ISSN

0743-7463

Publication Date

May 2009

Volume

25

Issue

10

Start / End Page

5725 / 5730

Related Subject Headings

  • Umbilical Veins
  • Streptavidin
  • Polytetrafluoroethylene
  • Oligopeptides
  • Ligands
  • Humans
  • Fibronectins
  • Endothelial Cells
  • Chemical Physics
  • Cells, Cultured
 

Citation

APA
Chicago
ICMJE
MLA
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Anamelechi, C. C., Clermont, E. C., Novak, M. T., & Reichert, W. M. (2009). Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin. Langmuir : The ACS Journal of Surfaces and Colloids, 25(10), 5725–5730. https://doi.org/10.1021/la803963r
Anamelechi, Charles C., Edward C. Clermont, Matthew T. Novak, and William M. Reichert. “Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin.Langmuir : The ACS Journal of Surfaces and Colloids 25, no. 10 (May 2009): 5725–30. https://doi.org/10.1021/la803963r.
Anamelechi CC, Clermont EC, Novak MT, Reichert WM. Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin. Langmuir : the ACS journal of surfaces and colloids. 2009 May;25(10):5725–30.
Anamelechi, Charles C., et al. “Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin.Langmuir : The ACS Journal of Surfaces and Colloids, vol. 25, no. 10, May 2009, pp. 5725–30. Epmc, doi:10.1021/la803963r.
Anamelechi CC, Clermont EC, Novak MT, Reichert WM. Dynamic seeding of perfusing human umbilical vein endothelial cells (HUVECs) onto dual-function cell adhesion ligands: Arg-Gly-Asp (RGD)-streptavidin and biotinylated fibronectin. Langmuir : the ACS journal of surfaces and colloids. 2009 May;25(10):5725–5730.
Journal cover image

Published In

Langmuir : the ACS journal of surfaces and colloids

DOI

EISSN

1520-5827

ISSN

0743-7463

Publication Date

May 2009

Volume

25

Issue

10

Start / End Page

5725 / 5730

Related Subject Headings

  • Umbilical Veins
  • Streptavidin
  • Polytetrafluoroethylene
  • Oligopeptides
  • Ligands
  • Humans
  • Fibronectins
  • Endothelial Cells
  • Chemical Physics
  • Cells, Cultured