Endothelial cell Seeding: Coating Dacron and Expanded Polytetrafluoroethylene Vascular Grafts with a Biological Glue Allows Adhesion and Growth of Human Saphenous Vein Endothelial cells

Mazzucotelli, J. P.; Klein-Soyer, C; Beretz, Alain; Brisson, Christine; Archipoff, G; Cazenave, Jean‐Pierre

doi:10.1177/039139889101400806

Cited by 40 publications

(18 citation statements)

References 41 publications

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“…35,36 After they were thawed, the cells were seeded on purified human FN in culture dishes and used from the second to the fifth passages. The culture medium was medium 199/RPMI 1640 (50/50) containing 10 mmol/L HEPES, 2 mmol/L L-glutamine, 100 U/mL penicillin, 100 g/mL streptomycin, 0.25 g/mL fungizone, and 30% pooled heat-inactivated HS.…”

Section: Cell Culturementioning

confidence: 99%

CD9 Participates in Endothelial Cell Migration During In Vitro Wound Repair

Klein-Soyer

Azorsa

Cazenave

et al. 2000

ATVB

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Abstract-CD9, a widely expressed membrane protein of the tetraspanin family, has been implicated in diverse functions, such as signal transduction, cell adhesion, and cell motility. We tested the effects of an anti-CD9 monoclonal antibody (ALMA.1) on the migration and proliferation of human vascular endothelial cells (ECs) during repair of an in vitro mechanical wound mimicking angiogenic processes. ALMA.1 induced dose-dependent inhibition of wound repair with a 35Ϯ1.5% decrease at 20 g/mL. Only cell migration was affected, because the rate of proliferation of ECs at the lesion margin was not modified and because the inhibition of repair was also observed for nonproliferating irradiated ECs. Monoclonal antibodies against CD63 tetraspanin (H5C6) and control mouse IgG (MOPC-21) were inactive. CD9, one of the most abundant proteins at the surface of ECs, colocalized with ␤ 1 or ␤ 3 integrins on EC membranes in double-labeling immunofluorescence experiments with ALMA.1 and an anti-␤ 1 (4B4) or anti-␤ 3 (SDF.3) monoclonal antibody. Moreover, ALMA.1 and 4B4 had additive inhibitory effects on lesion repair, whereas 4B4 alone also inhibited EC proliferation. In transmembrane Boyden-type assays, ALMA.1 induced dose-dependent inhibition of EC migration toward fibronectin and vitronectin with 45Ϯ6% and 31Ϯ10% inhibition, respectively, at 100 g/mL. 4B4 inhibited migration toward fibronectin at 10 g/mL but had no effect in the case of vitronectin. Adhesion of ECs to immobilized anti-CD9 monoclonal antibodies induced tyrosine-phosphorylated protein levels similar to those observed during interactions with ␤ 1 or ␤ 3 integrins. These results point to the involvement of CD9 in EC adhesion and migration during lesion repair and angiogenesis, probably through cooperation with integrins. As such, CD9 is a potential target to inhibit angiogenesis in metastatic and atherosclerotic processes.

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Section: Cell Culturementioning

confidence: 99%

CD9 Participates in Endothelial Cell Migration During In Vitro Wound Repair

Klein-Soyer

Azorsa

Cazenave

et al. 2000

ATVB

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“…[17][18][19][20] Our previous work 23 has shown the feasibility of introducing avidin-biotin for increasing cell attachment and also has confirmed the need for integrins to promote cellular spreading. The study used a homogeneous ligand system of biotinylated BAEC attached onto avidin-treated glass slides and confirmed the hypothesis that a high-affinity receptor ligand system could increase the strength of initial cell attachment.…”

Section: Discussionmentioning

confidence: 99%

“…Previous work by other researchers [17][18][19][20] using a combination of integrin-mediated serum protein treatments (Fn, fibrinogen, von Willebrand factor, etc. ), have shown improved cell adhesion compared to individual protein treatments.…”

Section: Discussionmentioning

confidence: 99%

“…Single-angle TIRFM permits relative quantification of the cell-substrate separation distances. BAEC adherent to the glass slides were labeled with 3 g/l 1,1Ј-dioctadecyl-3,3,3Ј,3Ј-tetramethyl-indocarbo-cyanineperchlorate (DiIC 18 3 , Molecular Probes, Eugene, OR) and observed using TIRFM, as described elsewhere. 34 The intrinsic error of the model used to determine the membrane/substratum separation was 10%.…”

Section: Total Internal Reflection Fluorescence Microscopy (Tirfm)mentioning

confidence: 99%

“…6,7,9,10,[14][15][16] A few studies have investigated a multiple ligand-based approach for the enhancement of EC adhesion and have shown that a combination of proteins support better EC adhesion than do single proteins. [17][18][19][20] However, cell adhesion through all these proteins utilized the integrin-mediated adhesion system. Recent reports have shown avidin-biotin mediates binding to convert nonadhesive mouse ascites carcinoma cells to anchorage-dependent cells.…”

Section: Introductionmentioning

confidence: 99%

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Fibronectin and avidin-biotin as a heterogeneous ligand system for enhanced endothelial cell adhesion

Bhat

Truskey

Reichert

1998

J. Biomed. Mater. Res.

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A preadsorbed layer of "heterogeneous" integrin-dependent and -independent protein was used to enhance initial integrin-mediated endothelial cell attachment and spreading. Glass substrates were treated with fibronectin (Fn) and avidin coupled through adsorbed biotinylated bovine serum albumin (b-BSA). The slides then were seeded with biotinylated BAEC. Control "homogeneous" surfaces were slides adsorbed with either Fn or avidin coupled to b-BSA. The cells were incubated for 0.5 h in serum-containing media and exposed to a range of shear stresses in a laminar flow variable-height flow chamber. The critical shear stress to detach 50% of the seeded cells on the heterogeneous ligand surface was significantly greater than for either of the control homogeneous ligand systems (p < 0.001). Cellular spreading during the initial period of 0-2 h also was higher (p < 0.05) on the heterogeneous ligand-treated surface than on the surface of either of the homogeneous controls. The close contact area of the cell membrane with the substrate 1 h after seeding in serum-containing media was measured using TIRFM. Cells attached onto the heterogeneous ligand-treated surfaces had a significantly (p < 0.01) higher area of close contact with the substrate, which is consistent with a greater degree of attachment and spreading. The results indicate that the combination of integrin-dependent and -independent adhesion systems using heterogeneous ligands further enhances initial endothelial cell attachment and spreading.

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Using avidin-mediated binding to enhance initial endothelial cell attachment and spreading

Bhat

Truskey

Reichert

1998

J. Biomed. Mater. Res.

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Binding between the protein avidin and the vitamin biotin was used as an extrinsic, high affinity receptor-ligand system to augment the intrinsic integrin-dependent cellular adhesion mechanism. Glass substrates were coupled with avidin receptors through an adsorbed film of biotinylated bovine serum albumin (b-BSA). The avidin-treated slides then were seeded with biotinylated bovine aortic endothelial cells (BAEC). A 3:1 ratio of BSA:b-BSA provided the best results in terms of specific cellular attachment, growth, and spreading. Control surfaces consisted of bare glass or glass with adsorbed BSA. Attachment of unmodified BAEC to glass decreased in the presence of anti-beta 1 integrin antibody. Adhesion of biotinylated BAEC to avidin-treated slides was not affected by anti-beta 1 integrin antibody, consistent with integrin-independent avidin-mediated adhesion. The initial rate of cell spreading was greatest for avidin-biotin-mediated adhesion (80.0 +/- 25.6 microns2/h), followed by integrin-dependent cellular adhesion on plain glass (35.7 +/- 7.7 microns2/h) and, finally, by adhesion on BSA-coated protein surfaces (10.2 +/- 0.3 microns2/h). Biotinylated and unmodified BAEC, cultured for 1 h in serum-containing media, were subjected to laminar flow in a variable-height flow chamber that provided a range of shear stresses from 0.2 to 75 dynes/cm2. The critical shear stress required to detach 50% of the cells in serum-containing media increased from 4.6 +/- 0.8 dynes/cm2 for integrin-dependent adhesion to 12.6 +/- 1.2 dynes/cm2 for avidin-biotin-mediated adhesion. Avidin-mediated attachment for biotinylated BAEC increased initial cellular spreading rates and strength of attachment (i.e., at 1 h) by a factor of two and three, respectively. These results support the hypothesis that integrin-mediated cell attachment and spreading can be enhanced using high affinity integrin-independent binding.

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Endothelial cell Seeding: Coating Dacron and Expanded Polytetrafluoroethylene Vascular Grafts with a Biological Glue Allows Adhesion and Growth of Human Saphenous Vein Endothelial cells

Cited by 40 publications

References 41 publications

CD9 Participates in Endothelial Cell Migration During In Vitro Wound Repair

CD9 Participates in Endothelial Cell Migration During In Vitro Wound Repair

Fibronectin and avidin-biotin as a heterogeneous ligand system for enhanced endothelial cell adhesion

Using avidin-mediated binding to enhance initial endothelial cell attachment and spreading

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