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Isolation and characterization of microvessel endothelial cells from human mammary adipose tissue

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Summary

A method for the isolation and long-term culture of human microvessel endothelial cells from mammary adipose tissue (HuMMEC) obtained at breast reduction surgery has been developed. Pure cultures of HuMMEC were isolated by sequential digestion of the fat with collagenase and trypsin followed by specific selection of microvessel fragments withUlex europaeus agglutinin-1 coated magnetic beads (Dynabeads). The resulting cells formed contact-inhibited monolayers on gelatin and fibronectin substrates and capillary-like “tubes” on Matrigel; they also expressed von Willebrand factor, angiotensin-converting enzyme, and accumulated acetylated low density lipoprotein. Further immunofluorescence characterization revealed the presence of antigens for the endothelial cell specific monoclonal antibodies EN4 and H4-7/33. In addition, the origin of these cells was confirmed by the demonstration of the cell adhesion molecules, platelet endothelial cell adhesion molecule-1 (CD31), and endothelial leukocyte adhesion molecule-1 (ELAM-1/E-selectin) upon stimulation with tumor necrosis factor (TNF)α. HuMMEC were found to express-1 ELAM-1 at lower levels of TNFα (<10 ng/ml) than required by human umbilical vein endothelial cells. These cells should provide a useful in vitro model for studying various aspects of microvascular biology and pathology.

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References

  • Anders, E.; Alles, J. U.; Delvos, U., et al. Microvascular endothelial cells from human omental tissue: modified method for long-term cultivation and new aspects of characterization. Microvasc. Res. 34:239–249; 1987.

    Article  PubMed  CAS  Google Scholar 

  • Auerbach, R.; Alby, L.; Grieves, J., et al. Monoclonal antibody against angiotensin-converting enzyme: its use as a marker for murine, bovine and human endothelial cells. Proc. Natl. Acad. Sci. USA 79:7891–7895; 1982.

    Article  PubMed  CAS  Google Scholar 

  • Auerbach, R.; Alby, L.; Morrissey, L. W., et al. Expression of organ-specific antigens on capillary endothelial cells. Microvasc. Res. 29:401–411; 1985.

    Article  PubMed  CAS  Google Scholar 

  • Belloni, P. N.; Nicholson, G. L. Differential of cell surface glycoproteins on various organ derived microvascular endothelia and endothelial cell cultures. J. Cell Physiol. 136:398–410; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Bloom, W.; Fawcett, D. W. A textbook of histology. Philadelphia: W. B. Saunders Co.; 1975:83–84.

    Google Scholar 

  • Carson, M. P.; Haudenschild, C. C. Microvascular endothelium and pericytes: high yield, low passage cultures. In Vitro Cell. Dev. Biol. 22:344–354; 1986.

    Article  PubMed  CAS  Google Scholar 

  • Chung-Welch, N.; Patton, W. F.; Yen-Patton, G. P. A., et al. Phenotypic comparison between mesothelial and microvascular endothelial cell lineages using conventional endothelial cell markers, cytoskeletal protein markers andin vitro assays of angiogenic potential. Differentiation 42:44–53; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Chung-Welch, N.; Shepro, D.; Dunham, B., et al. Prostacyclin and prostaglandin secretions by bovine pulmonary microvessel endothelial cells are altered by changes in culture conditions. J. Cell Physiol. 135:224–234; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Cui, Y. C.; Tai, P-C.; Gatter, K. C., et al. A vascular endothelial cell antigen with restricted distribution in human foetal adult and malignant tissues. Immunology 49:183–189; 1983.

    PubMed  CAS  Google Scholar 

  • Davison, P. M.; Bensch, K.; Karasek, M. A. Isolation and long-term serial cultivation of endothelial cells from the microvessels of the adult human dermis. In Vitro Cell. Dev. Biol. 19:937–945; 1983.

    CAS  Google Scholar 

  • Dorovini-Zis, K.; Prameya, R.; Bowman, P. D. Culture and characterization of microvessel endothelial cells derived from human brain. Lab. Invest. 64:425–436; 1991.

    PubMed  CAS  Google Scholar 

  • Drake, B. L.; Loke, Y. W. Isolation of endothelial cells from human first trimester decidua using immunomagnetic beads. Hum. Reprod (Oxf) 6:1156–1159; 1991.

    CAS  Google Scholar 

  • Folkman, J.; Haudenschild, C. C.; Zetter, B. R. Long-term culture of capillary endothelial cells. Proc. Natl. Acad. Sci. USA 76:5217–5221; 1979.

    Article  PubMed  CAS  Google Scholar 

  • Fukuda, K.; Imamura, Y.; Koshihara, Y., et al. Establishment of human mucosal microvascular endothelial cells from inferior turbinate in culture. Am. J. Otolaryngol 10:85–91; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Gallery, E. D. M.; Rowe, J.; Schrieber, L., et al. Isolation and purification of microvascular endothelium from human decidual tissue in the late phase of pregnancy. Am. J. Obstet. Gynecol. 165:191–196; 1991.

    PubMed  CAS  Google Scholar 

  • Gilbert, S. F.; Migeon, B. R.d-Valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5:11–17; 1975.

    Article  PubMed  CAS  Google Scholar 

  • Gimbrone, M. A.; Cotran, R. S.; Folkman, J. Human vascular endothelial cells in culture, growth and DNA synthesis. J. Cell Biol. 60:673–674; 1974.

    Article  PubMed  CAS  Google Scholar 

  • Harach, H. R.; Jasani, B.; Williams, E. D. Factor VIII as a marker of endothelial cells in follicular carcinoma of the thyroid. J. Clin. Pathol. 36:1050–1054; 1983.

    PubMed  CAS  Google Scholar 

  • Holthöfer, H.; Virtanen, I.; Kariniemi, A-L., et al.Ulex europaeus 1 lectin as a marker for vascular endothelium in human tissues. Lab. Invest. 47:60–66; 1982.

    PubMed  Google Scholar 

  • Hormia, M.; Lehto, V. P.; Virtanen, I. Identification of UEA-1 binding surface glycoproteins on cultured human endothelial cells. Cell Biol. Int. Rep. 7:467–475; 1983.

    Article  PubMed  CAS  Google Scholar 

  • Imicke, E.; Ruszczak, Z. B.; Mayer-Da Silva, A., et al. Cultivation of human dermal microvascular endothelial cellsin vitro: immunocytochemical and ultrastructural characterization and effect of treatment with synthetic retinoids. Arch. Dermatol. Res. 283:149–157; 1991.

    Article  Google Scholar 

  • Jackson, C. J.; Garbett, P. K.; Nissen, B., et al. Binding of human endothelium toUlex europaeus-1 coated dyna-beads: application to the isolation of microvascular endothelium. J. Cell Sci. 96:257–262; 1990.

    PubMed  Google Scholar 

  • Jaffe, E. A.; Nachman, R. L.; Becker, C. G., et al. Culture of human endothelial cells derived from umbilical veins: identification by morphological criteria. J. Clin. Invest. 52:2745–2756; 1973.

    PubMed  CAS  Google Scholar 

  • Joos, F. The cerebral microvessels in culture, an update. J. Neurochem. 58:1–17; 1992.

    Article  Google Scholar 

  • Karasek, M. A. Microvascular endothelial culture. J. Invest. Dermatol. 93:335–385; 1989.

    Article  Google Scholar 

  • Kern, P. A.; Knedler, A.; Eckel, K. H. Isolation and culture of microvascular endothelium from human adipose tissue. J. Clin. Invest. 71:1822–1829; 1983.

    Article  PubMed  CAS  Google Scholar 

  • Knedler, A.; Eckel, R. H.; Kern, P. A., et al. Microvascular endothelial cell cultures from human omental adipose tissue. In Vitro Cell. Dev. Biol. 25:863–864; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Kumar, S.; West, D. C.; Ager, M. Heterogeneity in endothelial cells from large vessels and microvessels. Differentiation 36:57–70; 1987.

    Article  PubMed  CAS  Google Scholar 

  • Kuzu, I.; Bicknell, R.; Harris, A. M., et al. Heterogeneity of vascular endothelial cells with relevance to diagnosis of vascular tumors. J. Clin. Pathol. 45:143–148; 1992.

    PubMed  CAS  Google Scholar 

  • Latron, Y.; Alessi, M. C.; George, F., et al. Characterization of epitheloid cells from human omentum: comparison with endothelial cells from umbilical veins. Thromb. Haemostasis 66:361–367; 1991.

    CAS  Google Scholar 

  • Lodge, P. A.; Haisch, G. E.; Huber, S. A., et al. Biological differences in endothelial cells depending upon organ differentiation. Trans. Proceedings 23:216–218; 1991.

    CAS  Google Scholar 

  • Marks, R. M.; Czerniecki, M.; Penny, R. Human dermal microvascular endothelial cells: an improved method for tissue culture and a description of some singular properties in culture. In Vitro Cell. Dev. Biol. 21:627–635; 1985.

    Article  PubMed  CAS  Google Scholar 

  • Miettinen, M.; Holthöffer, H.; Lehto, V-P., et al.Ulex europaeus lectin as a marker for tumors derived from endothelial cells. Am. J. Clin. Pathol. 79:32–36; 1983.

    PubMed  CAS  Google Scholar 

  • Newman, P. J.; Berndt, M. C.; Gorshi, J., et al. PECAM-1 (CD31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily. Science 247:1219–1222; 1990.

    Article  PubMed  CAS  Google Scholar 

  • Phillips, M. L.; Nudelman, E.; Gaeta, F. C. A., et al. ELAM-1 mediates cell adhesion by recognition of a carbohydrate ligand Sialyl-LeX. Science 250:1130–1132; 1990.

    Article  PubMed  CAS  Google Scholar 

  • Picciano, P. T.; Johnson, B.; Walenga, R. W. Effects ofd-valine on pulmonary artery endothelial cell morphology and cell function in cell culture. Cell 151:134–147; 1984.

    Article  CAS  Google Scholar 

  • Pötzsch, B.; Grulich-Henn, J.; Rössing, R., et al. Identification of endothelial and mesothelial cells in human omental tissue and in omentumderived cultured cells by specific cell markers. J. Lab. Invest. 63:841–852; 1990.

    Google Scholar 

  • Rupnick, M. A.; Carey, A.; Williams, S. K. Phenotypic diversity in cultured cerebral microvascular endothelial cells. In Vitro Cell. Dev. Biol. 24:435–444; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Satoh, K.; Prescott, S. M. Culture of mesothelial cells from bovine pericardium and characterization of their arachidonate metabolism. Bio-Chem. Biophys. Acta 930:283–296; 1987.

    Article  CAS  Google Scholar 

  • Simmons, D. L.; Walker, C.; Power, C., et al. Molecular cloning of cd31, a putative intercellular adhesion molecule closely related to carcinoembryonic antigen. J. Exp. Med. 171:2147–2153; 1990.

    Article  PubMed  CAS  Google Scholar 

  • Stoltz, D.; Jacobson, B. Macro- and microvascular endothelial cells in vitro: maintenance of biochemical heterogeneity despite loss of ultrastructural characteristics. In Vitro Cell. Dev. Biol. 27A:169–182; 1991.

    Google Scholar 

  • Stylianou, E.; Jenner, L. A.; Davies, M., et al. Isolation, culture and characterization of human peritoneal mesothelial cells. Kidney Int. 37:1563–1570; 1990.

    PubMed  CAS  Google Scholar 

  • Takahasi, K.; Hata, J-I.; Mukai, K., et al. Close similarity between cultured human omental mesothelial cells and endothelial cells in cytochemical markers and plasminogen activator production. In Vitro Cell. Dev. Biol. 27a:542–548; 1991.

    Google Scholar 

  • Takahashi, K.; Sawasaki, Y.; Goto, T., et al. Cobblestone monolayer cells from human omental adipose tissue are possibly mesothelial not endothelial. In Vitro Cell Dev. Biol. 25:109–111; 1989.

    Article  PubMed  CAS  Google Scholar 

  • van Hinsbergh, V. W. M.; Kooistra, T.; Scheffer, M. A., et al. Characterization and fibrinolytic properties of human omental tissue mesothelial cells: comparison with endothelial cells. Blood 75:1490–1497; 1990.

    PubMed  Google Scholar 

  • Visser, M. J. P.; van Bockel, J. J.; van Muijen, G. N. P., et al. Cells derived from omental fat tissue and used for seeding vascular prostheses are not endothelial in origin: a study on the origin of epitheloid cells derived from human omentum. J. Vasc. Surg. 13:373–381; 1991.

    Article  PubMed  CAS  Google Scholar 

  • Voyta, J. C.; Via, D. P.; Butterfield, C. E., et al. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J. Cell Biol. 99:2034–2040; 1984.

    Article  PubMed  CAS  Google Scholar 

  • Walker, R. A.Ulex europaeus 1-peroxidase as a marker of vascular endothelium: its application in routine histopathology. J. Pathol. 146:123–127; 1985.

    Article  PubMed  CAS  Google Scholar 

  • Wagner, R. C.; Matthews, M. A. The isolation and culture of capillary endothelium from epididymal fat. Microvasc. Res. 10:286–297; 1975.

    Article  PubMed  CAS  Google Scholar 

  • Watkins, M. T.; Sharefkin, J. B.; Zajtchuk, R., et al. Adult human saphenous vein endothelial cells: assessment of their reproductive capacity for use in endothelial seeding of vascular prostheses. J. Surg. Res. 36:588–596; 1984.

    Article  PubMed  CAS  Google Scholar 

  • Williams, S. K.; Jarrell, B. E.; Rose, D. G., et al. Human microvessel endothelial cell isolation and vascular graft sodding in the operating room. Annal. Vasc. Surg. 3:147–152; 1989.

    Google Scholar 

  • Wolff, J. E. A.; Belloni-Olivi, L.; Bressler, J. P., et al.γ-Glutamyltranspeptidase activity in brain microvessels exhibits regional heterogeneity. J. Neurochem. 58:909–915; 1992.

    Article  PubMed  CAS  Google Scholar 

  • Zetter, B. R. Endothelial heterogeneity: influence of vessel size, organ localization and species specificity on the properties of cultured endothelial cells. In: Ryan, U. S., ed. Endothelial cells, vol. 2. Boca Raton, FL: CRC Press; 1988.

    Google Scholar 

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  1. Endothelial Biology Group, CRC Gray Laboratory, Mount Vernon Hospital, P.O. Box 100, HA6 2JR, Northwood, Middlesex, United Kingdom

    P. W. Hewett, J. C. Murray, E. A. Price, M. E. Watts & M. Woodcock

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  1. P. W. Hewett
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  2. J. C. Murray
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  3. E. A. Price
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  4. M. E. Watts
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  5. M. Woodcock
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Hewett, P.W., Murray, J.C., Price, E.A. et al. Isolation and characterization of microvessel endothelial cells from human mammary adipose tissue. In Vitro Cell Dev Biol - Animal 29, 325–331 (1993). https://doi.org/10.1007/BF02633961

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  • DOI: https://doi.org/10.1007/BF02633961

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