This paper is only available as a PDF. To read, Please Download here.
Abstract
Prelining graft material with autologous functioning endothelial cells might be one
of the ultimate requirements to obtain a biocompatible surface. Accordingly, endothelial
cells from stripped varicose veins were enzymatically harvested and grown on a fibronectin
matrix. Proliferation was investigated in defined medium supplemented with various
concentrations of endothelial cell growth supplement (ECGS) (25, up to 150 μg/ml)
and heparin (10−8, up to 10−5mol/L): optimal growth required both 150 μg/ml of ECGS and 10−5mol/L heparin. Under these conditions, cell culture achieved cell densities at a confluence
of 1.2 ± 1.1 105 cells/cm2 with a doubling time of 1 day. During subcultivation cultured cells consistently
exhibited characteristic cobblestone morphology and immunofluorescent staining for
factor VIII-related antigen, whereas prostacyclin production determined by enzyme-linked
immunosorbent assay for 6-keto-prostaglandin F1α reached 21.1 ± 1.2 ng/106 cells after 15-minute stimulation with 1 U/ml of thrombin. Heparin-containing culture
medium-endothelial cell interactions were particularly studied, and with iodine 125-heparin,
binding was demonstrated with an apparent dissociation constant (Kd) of 0.36 ± 0.04
μmol/L. A cold storage technique at −80 °C was sought, and freezed cells were used
to coat in vitro polytetrafluoroethylene grafts. Protein-treated material allowed
cell attachment and growth to a confluent monolayer as assayed by light and scanning
electron microscopy. These data validate the feasibility of prelining grafts in vitro
with autologous functioning endothelial cells. This approach may be useful in improving
the performance of small-caliber vascular grafts according to prostacyclin production
and surface-bound heparin of these cells.
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to SurgeryAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Presidential address: the microcosm of the arterial wall. A plea for research.J Vasc Surg. 1987; 5: 1-18
- Expanded polytetrafluoroethylene vascular prosthesis seeded with enzymatically derived and cultured canine endothelial cells.Surgery. 1982; 91: 550-559
- Early normalization of platelet survival by endothelial seeding of Dacron arterial prostheses in dogs.Surgery. 1982; 92: 385-393
- Endothelial cell seeding of small-caliber synthetic grafts in the baboon.Surgery. 1986; 99: 318-325
- Adult human endothelial cell coverage of small-caliber Dacron and polytetrafluoroethylene vascular prosthesis in vitro.J Surg Res. 1986; 41: 158-172
- Use of an endothelial monolayer on a vascular graft prior to implantation.Ann Surg. 1986; 203: 671-678
- The preparation of 131I-labeled human growth hormone of high specific radioactivity.Biochem J. 1963; 89: 114-123
- Binding of heparin and low molecular weight heparin fragments to human vascular endothelial cells in culture.Nouv Rev Fr Hematol. 1984; 26: 243-247
- What does I50 mean?.Biochem J. 1983; 215: 429-430
- A single-staged technique for seeding vascular grafts with autogenous endothelium.Surgery. 1978; 84: 498-504
- Influence of endothelial cell seeding on platelet deposition and patency in small-diameter Dacron arterial grafts.J Vasc Surg. 1984; 1: 224-233
- Endothelium develops on seeded human arterial prosthesis: a brief clinical note.J Vasc Surg. 1985; 2: 727-730
- Seeding human arterial prostheses with mechanically derived endothelium. The detrimental effect of smoking.J Vasc Surg. 1984; 1: 279-289
- Endothelial seeding of polytetrafluoroethylene popliteal bypasses: a preliminary report.J Vasc Surg. 1987; 6: 114-118
- Reduction in deposition of indium 111-labeled platelets after autologous endothelial cell seeding of Dacron aortic bifurcation grafts in humans: a preliminary report.J Vasc Surg. 1987; 6: 17-25
- Varicose veins as a source of adult human endothelium cells.Tissue Cell. 1985; 17: 171-176
- Comparative effects of trypsin, collagenase and mechanical harvesting on cell membrane lipids studied in monolayer-cultured endothelial cells and a green monkey kidney cell line.Biochim Biophys Acta. 1985; 846: 120-126
- Use of freshly isolated capillary endothelial cells for the immediate establishment of a monolayer on a vascular graft at surgery.Surgery. 1986; 100: 392-398
- Serial propagation of human endothelial cells in vitro.J Cell Biol. 1981; 91: 420-426
- Modulation of endothelial cell growth by heparin.Cell Biol Int Rep. 1986; 10: 437-446
- Human endothelial cells: use of heparin in cloning and long term serial cultivation.Science. 1983; 222: 623-625
- In vitro endothelialization of small-caliber vascular grafts.Surgery. 1987; 101: 577-586
- Initial adherence of human capillary endothelial cells to Dacron.J Surg Res. 1987; 42: 133-140
- The effect of extracellular matrix on the detachment of human endothelial cells.J Cell Physiol. 1984; 121: 467-471
- Properties of cultured endothelium from adult human vessels.Arteriosclerosis. 1984; 4: 4-13
- Heparin regulates the collagen phenotype of vascular smooth muscle cells: induced synthesis of an MR 60 000 collagen.J Cell Biol. 1985; 100: 613-619
- Endothelial linings: the effect of serine protease inhibition.J Surg Res. 1987; 43: 10-13
- Regulation of cyclooxygenase activity in human vascular tissue.in: Proceedings of the 6th Intern Congress on Prostaglandins and Related Compounds. International Congress on Prostaglandins and Related Compounds, Florence1986: 96A
- Binding and endocytosis of heparin by human endothelial cells in culture.Biochem Biophys Acta. 1985; 845: 196-203
- Endothelial binding sites for heparin: specificity and role in heparin neutralization.Biochem J. 1986; 238: 847-854
- Use of endothelium cultured on microcarriers as a model for the microcirculation.Lab Invest. 1982; 47: 498-504
- The observation of heparin on endothelium after injection.Thromb Res. 1976; 8: 195-204
- Binding of heparin on the surface of cultured human endothelial cells.Thromb Res. 1978; 12: 773-784
- Heparin, a misunderstood drug.Trends Pharmacol Sci. 1982; 3: 289-291
- Role of heparin and heparin like molecules in thrombosis and atherosclerosis.in: 6th ed. Fed Proc. 44. 1985: 404-409
- Inhibition of rat arterial smooth muscle cell proliferation by heparin. II. In vitro studies.Circ Res. 1980; 47: 578-583
- Inhibition of vascular smooth muscle cell migration by heparin like glycosaminoglycans.J Cell Physiol. 1984; 118: 253-256
- Mast cell heparin stimulates migration of capillary endothelial cells in vitro.J Exp Med. 1980; 152: 931-944
- Pharmacological studies on frozen stored canine saphenous veins and basilar arteries.Arch Pharmacol. 1986; 332: 74-78
- Seeding of arteriovenous prostheses with homologous endothelium. A preliminary report.J Vasc Surg. 1986; 3: 860-866
Article info
Publication history
Accepted:
April 6,
1988
Identification
Copyright
© 1989 Published by Elsevier Inc.
