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DOI: 10.1055/s-0038-1646421
Characterization of Epitheloid Cells from Human Omentum: Comparison with Endothelial Cells from Umbilical Veins
Publication History
Received 10 July 1990
Accepted 11 March 1991
Publication Date:
25 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199103/lookinside/thumbnails/10-1055-s-0038-1646421-1.jpg)
Summary
Capillary cells represent 95% of the vascular bed, and cells from large and micro-vessels do not express identical functions. In order to study the hormonal regulation of plasminogen activator inhibitor 1 (PAI-1) secretion by human capillary cells we used epithelial cells from omental tissue (HOTMEC). As their endothelial origin is subject to controversy, we attempted to determine their characteristics by comparing them to human umbilical vein endothelial cells (HUVEC). Morphological and biological criteria were studied. By phase contrast microscopy HOTMEC elicited a cobblestone pattern similar to HUVEC. Weibel-Palade bodies were not found in the cytoplasm with electron microscopy. Fluorescence microscopy studies indicated that HOTMEC took up acetylated-LDL more intensely than HUVEC, and showed no staining for von Willebrand factor. The phenotype of HOTMEC was studied by flow cytometry using monoclonal antibodies (mo Ab) directed against epitopes either specific for endothelial cells or for mesothelial cells. We showed that in our preparations only 10% of cells reacted with mo Ab specific for endothelial cells. About 60% of the HOTMEC were labelled with an antibody directed against mesothelial cells. HOTMEC expressed fibrinolytic factors. Tissue plasminogen activator (t-PA) levels in HOTMEC conditioned medium were 50 fold higher than those of HUVEC, and the PAI-1 secretions were identical in both cell types. Insulin which is known to increase PAI-1 synthesis by hepatocytes did not enhance the PAI-1 level either in HOTMEC or in HUVEC conditioned media. Our results suggested that morphological and functional methods did not allow discrimination between the cell types present in the omentum tissue. They also showed that the population obtained from the omental tissue by collagenase digestion is heterogeneous, with few cells expressing endothelial markers.
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References
- 1 Hamsten A, De Faire U, Walldius G, Dehlen G, Szamosi A, Landon C, Blombôck M, Wiman B. Plasminogen activator inhibitor in plasma: Risk for recurrent myocardial infarction. Lancet 1987; 2: 3-5
- 2 Paramo JA, Alfaro MJ, Rocha E. Post-operative changes in the plasmatic levels of tissue-type plasminogen activator and its fast-acting inhibitor. Relationship to deep vein thrombosis and influence of prophylaxis. Thromb Haemostas 1985; 54: 713-6
- 3 Van Mourik JA, Lawrence DA, Loskutoff DJ. Purification of an inhibitor of plasminogen activator (anti-activator) synthetized by endothelial cells. J Biol Chem 1984; 259: 14914-21
- 4 Philips M, Juul AG, Thorsen S. Human endothelial cells produce a plasminogen activator inhibitor and a tissue type plasminogen activator inhibitor complex. Biochim Biophys Acta 1984; 802: 99-110
- 5 Sprengers ED, Princen HMG, Kooistra T, Van Hinsbergh VWM. Inhibition of plasminogen activators by conditioned medium of human hepatocytes and hepatoma cell line Hep G2. J Lab Clin Med 1985; 105: 751-8
- 6 Erickson LA, Ginsberg MH, Loskutoff DJ. Detection and partial characterization of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-7
- 7 Vague P, Juhan-Vague I, Aillaud MF, Badier C, Viard R, Alessi MC, Collen D. Correlation between blood fibrinolytic activity, plasminogen activator inhibitor level, plasma insulin level, and relative body weight in normal and obese subjects. Metabolism 1986; 2: 250-3
- 8 Juhan-Vague I, Roul C, Alessi MC, Ardissone JP, Heim M, Vague P. Increased plasminogen activator inhibitor activity in non insulin dependent diabetic patients. Relationship with plasma insulin. Thromb Haemostas 1989; 61: 370-3
- 9 Juhan-Vague I, Alessi MC, Joly P, Thirion X, Vague P, Declerck PJ, Serradimigni A, Collen D. Plasma plasminogen activator inhibitor 1 in angina pectoris. Influence of plasma insulin and acute phase response. Arteriosclerosis 1989; 9: 362-7
- 10 Alessi MC, Juhan-Vague I, Kooistra T, Declerck PJ, Collen D. Insulin stimulates the synthesis of plasminogen activator inhibitor 1 by the human hepatocellular cell line Hep G2. Thromb Haemostas 1988; 60: 491-4
- 11 Kooistra T, Bosna PJ, Töns HAM, Van den Berg AP, Meyer P, Princen HMG. Plasminogen activator inhibitor 1: Biosynthesis and mRNA level are increased by insulin in cultured human hepatocytes. Thromb Haemostas 1989; 62: 723-8
- 12 Bar RS. Interaction of insulin and insulin-like growth factor (IgF) with endothelial cells. Am NY Acad Sci 1982; 401: 150-6
- 13 King GL, Buzney SM, Kahn CR, Hetu N, Buchwald S, McDonald SG, Rand LI. Differential responsiveness to insulin of endothelial and support cells from micro and macrovessels. J Clin Invest 1983; 71: 974-80
- 14 Kern PA, Knedler A, Allen LA. Isolation and culture of microvascular endothelium from human adipose tissue. J Clin Invest 1983; 71: 1822-9
- 15 Takahashi K, Goto T, Mukai K, Sawasaki Y, Hata J. Cobblestone monolayer cells from human omental adipose tissue are possibly mesothelial, not endothelial. In Vitro 1989; 25: 109-10
- 16 Van Hinsbergh V, Kooistra T, Scheffer M, Van Bockel M, Van Muijen G. Characterization and fibrinolytic properties of human omental tissue mesothelial cells. Comparison with endothelial cells. Blood 1990; 75: 1490-7
- 17 Poncelet P. Non lineage/natural killer section report: New and previously defined clusters. In: Leucocyte Typing IV. White Cell Differentiation Antigens.. Knapp W, Dörken B, Gilks WR, Rieber EP, Schmidt RE, Stein H, Von dem Borne AEG Kr. (eds) Oxford University Press: 1989. pp 541
- 18 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-56
- 19 Wagner DD, Olmstead JB, Marder JW. Immunolocalization of von Willebrad protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 1982; 95: 355-60
- 20 Pitas RE, Innerarity TL, Weinstein JN, Mahley RW. Acetoacetylated lipoproteins used to distinguish fibroblasts from macrophages in vitro by fluorescence microscopy. Arteriosclerosis 1981; 1: 177-85
- 21 Redgrave TG, Roberts CDK, West CE. Separation of plasma lipoproteins by density gradient ultracentrifugation. Anal Biochem 1975; 65: 42-6
- 22 Declerck PJ, Alessi MC, Vestreken M, Kruithof EKO, Juhan-Vague I, Collen D. Measurements of plasminogen activator inhibitor 1 (PAI-1) in biological fluids with a murine monoclonal antibody based enzyme linked immunosorbent assay. Blood 1988; 71: 220-5
- 23 Bergsdorf N, Nilsson T, Wallén P. An enzyme linked immunosorbent assay for determination of tissue plasminogen activator applied to patients with thromboembolic disease. Thromb Haemostas 1983; 50: 740-4
- 24 Verheijen JH, Chang GTG, Kluft C. Evidence for the occurrence of a fast acting inhibitor for tissue plasminogen activator in human plasma. Thromb Haemostas 1984; 51: 392-5
- 25 Hammersen F, Hammersen E. Some structural and functional aspects of endothelial cells. Basic Res Cardiol 1985; 80: 491-501
- 26 Antonov AS, Kikolaeva MA, Klueva TS, Romanov YA, Babacu VR, Bystrevskaya VB, Perov NA, Regin VS, Smirnov VN. Primary culture of endothelial cells from atherosclerotic human aorta. Atherosclerosis 1986; 59: 1-19
- 27 Rone JD, Goodman AL. Heterogeneity of rabbit aortic endothelial cells in primary cultures. Proc Soc Exp Biol Med 1985; 179: 136-40
- 28 Drouet L, Berard M, Peyri N, Baudin B, Carrier JL, Boffa MC. Are human omental micro-vascular endothelial cells (HOTMEC) really endothelial cells (EC) or mesothelial cells (MC). Thromb Haemostas 1989; 62: 578 (Abstr 1853)
- 29 Voyta J, Via D, Butterfield C, Zetter B. Identification and isolation of endothelial cells based on their increased uptake of acetylated low density lipoprotein. J Cell Biol 1984; 99: 2034-40
- 30 Havekes L, Mommaas-Kienhuis AM, Schouten D, De Wit E, Scheffer M, Van Hinsbergh VWM. High-affinity uptake and degradation of acetylated low density lipoprotein by confluent human vascular endothelial cells. Atherosclerosis 1985; 56: 81-92
- 31 Mahley RW, Innerarity T, Weisgraber K, Oh S. Altered metabolism (in vivo and in vitro) of plasma lipoproteins after selective chemical modification of lysine residues of the lipoprotein. J Clin Invest 1979; 64: 743-50
- 32 Goldstein JL, Ho YK, Basu SK, Brown MS. Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition. Proc Natl Acad Sci 1979; 76: 333-7
- 33 Leewenberg J, Jeunhomme TM, Buurman WA. Induction of an activation antigen on human endothelial cells in vitro. Eur J Immunol 1989; 19: 715-20
- 34 Dupuy E, Bikfalvi A, Rendu F, Levy Toledano S, Tobelem G. Thrombin mitogenic responses and protein phosphorylation are different in cultured human endothelial cells derived from large and microvessels. Exp Cell Res 1989; 185: 363-72
- 35 Jahn L, Fouquet B, Roke K, Franke WW. Cytokeratins in certain endothelial and smooth muscle cells of two toxonomically distant vertebrate species, Xenopus laevis and man. Differentiation 1987; 36: 234-54
- 36 Speiser W, Anders E, Preissner RT, Wagner O, Müller-Berghaus G. Differences in coagulant and fibrinolytic activities of cultured human endothelial cells derived from omental tissue microvessels and umbilical veins. Blood 1987; 69: 964-9
- 37 Kolodie L, Rachail M. Different molecular weight found of tissue-type plasminogen activator in ascitic fluid from patients with liver cirrhoses. Fibrinolysis 1988; 2: 245-9
- 38 Gerritsen ME, Burke TM, Allen LA. Glucose starvation is required for insulin stimulation of glucose uptake and metabolism in cultured microvascular endothelial cells. Microvasc Res 1988; 35: 153-66