Download PDF
Thromb Haemost 1995; 74(04): 1096-1102
DOI: 10.1055/s-0038-1649887
Original Article
Coagulation
Schattauer GmbH Stuttgart

Tissue Factor Expression on Mesothelial Cells is Induced during in vitro Culture – Manipulation of Culture Conditions Creates Perspectives for Mesothelial Cells as a Source for Cell Seeding Procedures on Vascular Grafts

Hence J M Verhagen
1   The Department of Surgery, University Hospital Utrecht, The Netherlands
2   The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Glenda J Heijnen-Snyder
2   The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Tom Vink
2   The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Apollo Pronk
1   The Department of Surgery, University Hospital Utrecht, The Netherlands
,
Theo J M V van Vroonhoven
1   The Department of Surgery, University Hospital Utrecht, The Netherlands
,
Bert C Eikelboom
1   The Department of Surgery, University Hospital Utrecht, The Netherlands
,
Jan J Sixma
2   The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Philip G de Groot
2   The Department of Haematology, University Hospital Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 16 January 1994

Accepted after resubmission 23 June 1995

Publication Date:
27 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Lining the luminal surface of prosthetic small diameter bypasses with endothelial cells (EC) will lower its thrombogenicity. Unfortunately, human EC are only scarcely available. Mesotheliai cells (MC) have antithrombotic properties in vivo and can be harvested in large numbers, from the omentum. Recent work demonstrated that the expression of tissue factor (TF) is induced in MC after isolation and culture. Different culture conditions were studied to suppress TF-expression.

MC grown in pooled human serum (HS) are procoagulant (717 ± 119 pM factor Xa/min.105 cells). Replacing HS for fetal calf serum, precoating the surface with extracellular matrix and the addition of the xanthine-oxidase inhibitor allopurinol, inhibited TF expression by 90% (p <0.001). Allopurinol clearly reduced TF-mRNA levels.

TF expression on cultured MC is an in-vitro effect due to culture conditions and the formation of oxygen free radicals. By reducing TF expression by 90%, we have established conditions in which MC are a good alternative for EC for seeding on synthetic grafts.

 

  • References

  • 1 Veith FJ, Gupta SK, Ascer E, White-Flores S, Samson RH, Scher LA, Towne JB, Bernhard VM, Bonier P, Flinn WR, Astelford P, Yao JS T, Bergan JJ. Six year prospective multicentre randomized comparison of autologous saphenous vein and expanded polytetrafluoroethylene grafts in infrainguinal arterial reconstructions. J Vase Surg 1986; 3: 104-114
    CrossrefPubMedGoogle Scholar
  • 2 Esquivel CO, Blaisdell FW. Why small caliber vascular grafts fail: a review of clinical and experimental experience and the significance of the interaction of the blood at the interface. J Surg Res 1986; 41: 1-5
    CrossrefPubMedGoogle Scholar
  • 3 Michaels JA. Choice of malerials for above-knee femoralpopliteal bypass graft. Br J Surg 1989; 76: 7-14
    CrossrefPubMedGoogle Scholar
  • 4 Oblath RW, Buckley RO, Green KM, Swartz SI, De Weese JA. Prevention of platelet aggregation and adherence to prosthetic vascular grafts by aspirin and dipyridamole. Surgery 1978; 84: 37-44
    PubMedGoogle Scholar
  • 5 Szilagyi DE. Vascular substitutes 1981. Achievements, disappointment, prospects J Cardiovasc Surg (Torino) 1982; 23: 183-193
    PubMedGoogle Scholar
  • 6 Kempczinski RF, Rosenman JE, Pearce WH, Rocdersheimcr LR, Berlatzky Y, Ramalanjaona GR. Endothelial cell seeding of a new PTEE vascular prosthescs. J Vase Surg 1985; 2: 424-429
    CrossrefPubMedGoogle Scholar
  • 7 Herring M, Dilley R, Cullison T, Gardner A, Glover J. Seeding endothelium on canine arterial prosthescs. The size of the inoculum J Surg Res 1980; 28: 35-38
    CrossrefPubMedGoogle Scholar
  • 8 Clarke JM F, Pittilo RM, Machin SJ, Woolf N. A study of the possible role of mcsothclium as a surface for flowing blood. Thromb Haemost 1984; 51: 57-60
    PubMedGoogle Scholar
  • 9 Bull HA, Pittilo RM, Drury J, Pollock JG, Clarke JM F, Woolf N, Marston A, Machin SJ. Effects of autologous mcsothelial seeding on prostacyclin production within Dacron arterial prosthescs. Br J Surg 1988; 75: 671-674
    CrossrefPubMedGoogle Scholar
  • 10 Thompson JN, Paterson-Brown S, Harbourne T, Whawell SA, Kalodiki E, Dudley HA F. Reduced human peritoneal plasminogen activating activity: possible mechanism of adhesion formation. Br J Surg 1989; 76: 382-384
    CrossrefPubMedGoogle Scholar
  • 11 Whitaker D, Papadimitriou JM, Walters M. The mesothelium and its reactions: a review. Crit Rev Toxicol 1982; 10: 81-144
    PubMedGoogle Scholar
  • 12 Pronk A, de Groot Ph G, Hoynck van Papendrccht AA G M, Verbrugh HA, Leguit P, van Vroonhoven Th J M V, Sixma JJ. Thrombogcnicity and procoagulant activity of human mcsothelial cells. Artcrioseler Thromb 1992; 12: 1428-1436
    PubMedGoogle Scholar
  • 13 Zur M, Nemerson Y. Tissue factor pathways of blood coagulation. In: Haemostasis and Thrombosis. Bloom AL, Thomas DP. eds. New York: Churchill Livingstone; 1987. pp 148-164
    Google Scholar
  • 14 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical cord veins: Identification of morphology and immunologic criteria. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 15 Willems C, Astaldi G, de Groot Ph G, Janssen MC, Gonsalvez MD, Zeijlemaker WP, van Mourik JA, van Aken WG. Media conditioned by cultured human vascular endothelial cells inhibit the growth of vascular smooth muscle cells. Exp Cell Res 1982; 139: 191-200
    CrossrefPubMedGoogle Scholar
  • 16 Houdijk WP M, de Groot Ph G, Nievelstein PF E M, Sakariassen KS, Sixma JJ. Subendothelial proteins and platelet adhesion: von Willebrand Factor and fibronectin, not thrombospondin, are involved in platelet adhesion to extracellular matrix of human vascular endothelial cells. Arteriosclerosis 1986; 6: 24-33
    CrossrefPubMedGoogle Scholar
  • 17 Bajai SP, Rapaport SI, Brown SF. Isolation and characterization of human factor VII. J Biol Chem 1981; 256: 253-259
    PubMedGoogle Scholar
  • 18 Miletich JP, Jackson CM, Majerus PW. Properties of the factor Xa binding site on human platelets. J Biol Chem 1978; 253: 6908-6916
    PubMedGoogle Scholar
  • 19 Meijers JC M, Tijburg PN M, Bouma BN. Inhibition of human blood coagulation factor Xa by alpha-2-macroglobulin. Biochemistry 1987; 26: 5932-5937
    CrossrefPubMedGoogle Scholar
  • 20 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-159
    PubMedGoogle Scholar
  • 21 Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd edit Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1989
    Google Scholar
  • 22 Potgens AJ G, Lubsen NH, van Altena G, Schoenmakers JG G, Ruiter DJ, de Waal RM W. Measurement of tissue factor messenger RNA levels in human endothelial cells by a quantitative RT-PCR assay. Thromb Haemost 1994; 71: 208-213
    PubMedGoogle Scholar
  • 23 Del Maestro RF. The influence of oxygen derived free radicals on in vitro and in vivo model systems. Acta Univ Upsaliensis 1979; 340: 332
    PubMedGoogle Scholar
  • 24 Reilly PM, Schiller HJ, Bulkley GB. The pharmacologic approach to tissue injury mediated by free radicals and other reactive oxygen metabolites. Am J Surg 1991; 161: 488-503
    CrossrefPubMedGoogle Scholar
  • 25 Shatos MA, Doherty JM, Hoak JC. Alterations in human vascular endothelial cell function by oxygen free radicals. Arterioscler Thromb 1991; 11: 594-601
    CrossrefPubMedGoogle Scholar
  • 26 Zwier JL, Kuppusamy P, Lutty GA. Measurement of endothelial cell free radical generation: Evidence for a central mechanism of free radical injury in postischaemic tissues. Proc Natl Acad Sci USA 1988; 85: 4046-4050
    CrossrefPubMedGoogle Scholar
  • 27 McCord JM, Fridovich I. The reduction of cytochrome c by milk xanthine oxydase. J Biol Chem 1968; 243: 5753-5760
    PubMedGoogle Scholar
  • 28 McCord JM. Oxygen-derived free radicals in postischemic tissue injury. N Engl J Med 1985; 312: 159-163
    CrossrefPubMedGoogle Scholar
  • 29 Bennister JV, Bannister WH, Rotillio G. Aspects of the structure, function, and applications of superoxide dismutase. CRC Crit Rev Biochem 1987; 22: 111-180
    PubMedGoogle Scholar
  • 30 Forman HJ, Fisher AB. Antioxidant defences. In: Oxygen and living processes an interdisciplinary approach. Gilbert DL. ed. New York: Springer-Verlag; 1981: 235-249
    Google Scholar
  • 31 Pasquier C, Franzini E, Abedinzadeh Z, Kaouadji MN, Hakim J. Gamma and pulse radiolysis study of pentoxifylline, a methylxanthine. Int J Radiat Biol 1991; 60: 433-47
    CrossrefPubMedGoogle Scholar
  • 32 Halliwell B, Gutteridge JM C. Role of iron in oxygen radical reactions. Methods Enzymol 1984; 105: 47-56
    PubMedGoogle Scholar
  • 33 Brach MA, de Vos S, Arnold C, Gruss HJ, Mertelsmann R, Herrmann F. Leukotriene B4 transcriptionally activates interleukin-6 expression involving NK-chi B and NF-IL6. Eur J Immunol 1992; 22: 2705-2711
    CrossrefPubMedGoogle Scholar
  • 34 Bravenboer B, Kappelle AC, Hamers EP T, van Buren T, Erkelens DW, Gispen WH. Potential use of glutathione for the prevention and treatment of diabetic neuropathy in the streptozotocin-induced diabetic rat. Diabetologica 1992; 35: 813-817
    CrossrefPubMedGoogle Scholar
  • 35 Seamon KB, Padgett W, Daly JW. Forskolin: unique diterpine activator of adenylate cyclase in membranes and in intact cells. Proc Natl Acad Sci USA 1981; 78: 3363-3367
    CrossrefPubMedGoogle Scholar
  • 36 Crutchley DJ, Conanan LB, Toledo AW, Solomon DE, Que BG. Effects of prostacyclin analogues on human endothelial cell tissue factor expression. Arterioscler Thromb 1993; 13: 1082-1089
    CrossrefPubMedGoogle Scholar
  • 37 Zwaginga JJ, de Boer HC, IJseldijk MJ W, Kerkhof A, Muller Berghaus, Gruhlich J, Sixma JJ, de Groot Ph G. Thrombogenicity of vascular cells. Comparison between endothelial cells isolated from different sources and smooth muscle cells and fibroblasts Arterioscler Thromb 1990; 10: 437-448
    CrossrefPubMedGoogle Scholar
  • 38 Ishii H, Horie S, Kizaki K, Kazama M. Retinoic acid counteracts both the downregulation of thrombomodulin and the induction of tissue factor in cultured human endothelial cells exposed to tumor necrosis factor. Blood 1992; 80: 2556-2562
    PubMedGoogle Scholar
  • 39 De Groot Ph G, Reinders JH, Sixma JJ. Perturbation of human endothelial cells by thrombin or PMA changes the reactivity of their extracellular matrix towards platelets. J Cell Biol 1987; 104: 697-704
    CrossrefPubMedGoogle Scholar