Endothelial Cell Seeding on PTFE Vascular Prostheses Using a Standardized Seeding Technique

1989 ◽  
Vol 12 (4) ◽  
pp. 270-275 ◽  
Author(s):  
J. Gerlach ◽  
K.-M. Kreusel ◽  
H.H. Schauwecker ◽  
E.S. Bücherl
Keyword(s):  
Endothelial Cells ◽  
Rotation Speed ◽  
Vascular Grafts ◽  
Cell Number ◽  
Cell Seeding ◽  
Vascular Prostheses ◽  
Adhesion Rate ◽  
Endothelial Cell Seeding ◽  
Kinetics Of ◽  
Graft Diameter

A standardized method was developed for seeding endothelial cells (EC) in tubular vascular grafts. A rotational cell seeding device for tubular prostheses is presented and parameters influencing the kinetics of cell adhesion (rotation speed, graft diameter, cell suspension level, inoculated cell number) are reported. Seeding EC in 14 mm ID PTFE vascular grafts with rotation rate of 10 rph gave an adhesion rate of 80% in a homogeneous monolayer.

Endothelial Cell Seeding on Crosslinked Collagen: Effects of Crosslinking on Endothelial Cell Proliferation and Functional Parameters

2000 ◽  
Vol 84 (08) ◽  
pp. 325-331 ◽  
Author(s):  
M. J. B. Wissink ◽  
M. J. A. van Luyn ◽  
R. Beernink ◽  
F. Dijk ◽  
A. A. Poot ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Plasminogen Activator ◽  
Type I Collagen ◽  
Vascular Grafts ◽  
Small Diameter ◽  
Endothelial Cell Proliferation ◽  
Type I ◽  
Cell Seeding ◽  
Endothelial Cell Seeding

SummaryEndothelial cell seeding, a promising method to improve the performance of small-diameter vascular grafts, requires a suitable substrate, such as crosslinked collagen. Commonly used crosslinking agents such as glutaraldehyde and formaldehyde cause, however, cytotoxic reactions and thereby hamper endothelialization of currently available collagen-coated vascular graft materials.The aim of this study was to investigate the effects of an alternative method for crosslinking of collagen, using N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC) in combination with N-hydroxysuccinimide (NHS), on various cellular functions of human umbilical vein endothelial cells (HUVECs) in vitro. Compared to non-crosslinked type I collagen, proliferation of seeded endothelial cells was significantly increased on EDC/NHS-crosslinked collagen. Furthermore, higher cell numbers were found with increasing crosslink densities. Neither the morphology of the cells nor the secretion of prostacyclin (PGI2), von Willebrand factor (vWF), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) was affected by the crosslink density of the collagen substrate. Therefore, EDC/NHScrosslinked collagen is candidate substrate for in vivo application such as endothelial cell seeding of collagen-coated vascular grafts.

Electrostatic Endothelial Cell Seeding Technique for Small-Diameter (<6 MM) Vascular Prostheses: Feasibility Testing

1997 ◽  
Vol 6 (6) ◽  
pp. 623-629 ◽  
Author(s):  
Gary L. Bowlin ◽  
Stanley E. Rittgers
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Surface Charge ◽  
Feasibility Study ◽  
Small Diameter ◽  
Cell Seeding ◽  
Vascular Prostheses ◽  
Endothelial Cell Seeding ◽  
Endothelial Cell Adhesion

Multiple studies have indicated the importance of surface charge in the adhesion of multiple cardiovascular cell lines including platelets and endothelial cells on the substrate materials (1,4,7-10,12-15). It is the purpose of this article to report a feasibility study conducted using an electrostatic endothelial cell seeding technique. The feasibility study was conducted using human umbilical vein endothelial cells (HUVEC), a static pool apparatus, a voltage source, and a parallel plate capacitor. The HUVEC concentration and seeding times were constant at 560,000 HUVEC/ml and 30 min, respectively. Scanning electron microscopy examination of the endothelial cell adhesion indicated that an induced temporary positive surface charge on e-PTFE graft material enhances the number and the maturation (flattening) of HUVECs adhered. The results indicated that the total number of endothelial cells adhered (70.9 mm2) was increased from 9198 ± 1194 HUVECs on the control (no induced surface charge) e-PTFE to 22,482 ± 4814 HUVECs (2.4 × control) on the maximum induced positive surface charge. The total number of cells in the flattened phase of adhesion increased from 837 ± 275 to 6785 ± 1012 HUVECs (8.1 ×) under identical conditions. Thus, the results of the feasibility study support the premise that electrostatic interaction is an important factor in both the endothelial cell adhesion and spreading processes and suggest that the electrostatic seeding technique may lead to an increased patency of small diameter (<6 mm) vascular prostheses.

A New, Automated and Accurate In Vitro Method to Quantify Endothelial Cells Attached to Vascular Prostheses

1994 ◽  
Vol 72 (01) ◽  
pp. 146-150 ◽  
Author(s):  
Michel J T Visser ◽  
Alexander C D van Lennep ◽  
J Hajo van Bockel ◽  
Victor W M van Hinsbergh ◽  
Jan Slats ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Functional Properties ◽  
Cell Seeding ◽  
Green Fluorescence ◽  
Vascular Prostheses ◽  
Endothelial Cell Seeding ◽  
Triton X ◽  
Accurate Quantification

SummaryOver a decade ago the idea of endothelial cell seeding was introduced in an attempt to improve the function of small caliber vascular prostheses. Although endothelial cell seeding is currently being applied clinically, several questions regarding the functional properties of the seeded endothelial cells remain. Evaluation of functional properties of endothelial cells on various types of vascular prostheses can be performed partly in vitro, but it is hampered by the fact that commonly used methods to quantify endothelial cells do not adequately apply to these cells on prosthetic materials.An accurate quantification method is described that is rapidly and easily applicable to endothelial cells attached to vascular prostheses. The method can also be used to quantify endothelial cells attached to culture dishes or microcarriers. Colorless, non-fluorescing, fluorescein-di-acetate was used, which was taken up by the attached endothelial cells, and which was then intracellularly converted to yellow fluorescein, emitting green fluorescence. Subsequently, triton-X-100 was appli-cated to release fluorescein and levels of fluorescence were measured with the automated aperture-defined microvolume (ADM) method, using an inverted fluorescence microscope to which a photometer was connected. The measured level of fluorescence is linearly related to endothelial cell numbers attached to prostheses. The accuracy and the reproducibility of cell countings are high.

Endothelial cell seeding of vascular grafts: Experimental investigations and clinical update

1996 ◽  
Vol 5 (5) ◽  
pp. 296 ◽  
Author(s):  
Manfred Deutsch ◽  
Johann Meinhart ◽  
Teddy Fischlein ◽  
Peter Zilla
Keyword(s):  
Endothelial Cell ◽  
Vascular Grafts ◽  
Experimental Investigations ◽  
Cell Seeding ◽  
Endothelial Cell Seeding

Endothelial Cell Seeding of Polytetrafluoroethylene Vascular Prostheses Coated With a Fibroblastic Matrix

1993 ◽  
Vol 7 (6) ◽  
pp. 549-555 ◽  
Author(s):  
J.M. Bellón ◽  
J. Buján ◽  
N.G. Honduvilla ◽  
A. Hernando ◽  
J. Navlet
Keyword(s):  
Endothelial Cell ◽  
Cell Seeding ◽  
Vascular Prostheses ◽  
Endothelial Cell Seeding

Microvascular Endothelial Cell Seeding of Small-Diameter Dacron|2TM|0 Vascular Grafts

1988 ◽  
Vol 1 (1) ◽  
pp. 35-44 ◽  
Author(s):  
Steven P. Schmidt ◽  
Navid Monajjem ◽  
M. Michelle Evancho ◽  
Todd R. Pippert ◽  
W. V. Sharp
Keyword(s):  
Endothelial Cell ◽  
Vascular Grafts ◽  
Small Diameter ◽  
Microvascular Endothelial Cell ◽  
Cell Seeding ◽  
Endothelial Cell Seeding ◽  
Microvascular Endothelial

Fibrinolytic profiles after endothelial cell seeding of prosthetic vascular grafts

1994 ◽  
Vol 8 ◽  
pp. 139
Author(s):  
J.V. Smyth ◽  
M. Welch ◽  
H.M.H. Carr ◽  
P. Eisenberg ◽  
M.G. Walker
Keyword(s):  
Endothelial Cell ◽  
Vascular Grafts ◽  
Cell Seeding ◽  
Endothelial Cell Seeding

Kinetics of endothelial cell seeding

1985 ◽  
Vol 2 (6) ◽  
pp. 778-784 ◽  
Author(s):  
J. E. Rosenman ◽  
R. F. Kempczinski ◽  
W. H. Pearce ◽  
Edward B. Silberstein
Keyword(s):  
Endothelial Cell ◽  
Cell Seeding ◽  
Endothelial Cell Seeding ◽  
Kinetics Of
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