Abstract:
Murine endothelial cells (ECs) have proven difficult to obtain and maintain in culture. Long-term maintenance of normal ECs remains a difficult task. In this article we report the establishment of the first cellular line of renal microvascular endothelium obtained from normal tissue. Cells were isolated, cloned, and maintained by serial passages for longer than 24 mo, using endothelial cell growth supplement (ECGS) and gelatin-coated plates. Their morphology and ultrastructure, expression of von Willebrand factor, presence of smooth muscle α-actin, vimentin, cytokeratin filaments, capillary structures formed on Matrigel, and some typical ECs surface molecules were the criteria used to characterize cultured ECs. When examined for responsiveness to Shiga toxin-1, 13-20% of cytotoxicity was observed when coincubated with lipopolysaccharides. This cytotoxicity was not observed for normal lung ECs (1G11). Consequently, REC-A4 line retains characteristics of resting microvascular ECs and represents a useful in vitro model to study biological and physiopathological properties of renal endothelium.
Registro:
Documento: |
Artículo
|
Título: | Isolation and molecular characterization of a mouse renal microvascular endothelial cell line |
Autor: | Gazzaniga, S.; González, L.; Mantovani, A.; Vecchi, A.; Wainstok, R. |
Filiación: | Departamento de Quimica Biologica, Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina Ist. di Ric. Farmacol. Mario Negri, Milan, Italy Departamento de Quimica Biologica, Fac. de Ciencias Exactas y Naturales, Ciudad Universitaria, 1428 Nuñez, Buenos Aires, Argentina
|
Palabras clave: | CD31; Endothelium; ICAM; Shiga toxin; VCAM; alpha actin; cytokeratin; gelatin; matrigel; vimentin; von Willebrand factor; animal cell; article; capillary; cell clone; cell culture; cell growth; cell isolation; cell surface; cell ultrastructure; controlled study; cytotoxicity; endothelium cell; in vitro study; kidney blood vessel; long term care; lung alveolus cell; material coating; microvasculature; molecular biology; morphological trait; nonhuman; pathophysiology; protein expression; vascular endothelium; Actins; Animals; Cell Line; Cell Separation; Cell Survival; Cercopithecus aethiops; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fluorescent Antibody Technique, Indirect; Keratins; Kidney; Lipopolysaccharides; Lipoproteins, LDL; Mice; Mice, Inbred BALB C; Serial Passage; Shiga-Like Toxin I; Vero Cells; Vimentin; von Willebrand Factor; Animalia; Murinae |
Año: | 2004
|
Volumen: | 40
|
Número: | 3-4
|
Página de inicio: | 82
|
Página de fin: | 88
|
DOI: |
http://dx.doi.org/10.1290/1543-706X(2004)040<0082:IAMCOA>2.0.CO;2 |
Título revista: | In Vitro Cellular and Developmental Biology - Animal
|
Título revista abreviado: | In Vitro Cell. Dev. Biol. Anim.
|
ISSN: | 10712690
|
CODEN: | ICDBE
|
CAS: | gelatin, 9000-70-8; matrigel, 119978-18-6; von Willebrand factor, 109319-16-6; acetyl-LDL; Actins; Keratins, 68238-35-7; Lipopolysaccharides; Lipoproteins, LDL; Shiga-Like Toxin I; Vimentin; von Willebrand Factor
|
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10712690_v40_n3-4_p82_Gazzaniga |
Referencias:
- Boegehold, M.A., Heterogeneity of endothelial function within the circulation (1998) Curr. Opin. Nephrol. Hypertens., 7 (1), pp. 71-78
- Craig, L.E., Spelman, J.P., Strandberg, J.D., Zink, M.C., Endothelial cells from diverse tissues exhibit differences in growth and morphology (1998) Microvasc. Res., 55, pp. 65-76
- Dong, Q.G., Bernasconi, S., Ostaglio, S.L., Wainstok De Calmanovici, R., Martín Padura, J., Brevario, F., Mantovani, A., Vecchi, A., A general strategy for isolation of endothelial cells from murine tissues: Characterization of two endothelial cell lines obtained from murine lung and subcutaneous sponge implants (1997) Arterioscler. Thromb. Vasc. Biol., 17 (8), pp. 1599-1604
- Fernández, G.C., Rubel, C., Dran, G., Gómez, S., Isturiz, M.A., Palermo, M.S., Shiga toxin-2 induces neutrophilia and neutrophil activation in a murine model of hemolytic uremic syndrome (2000) Clin. Immunol., 95 (3), pp. 227-234
- Folkman, J., Haundenschild, C.C., Zetter, B.R., Long-term culture of capillary endothelial cells (1979) Proc. Natl. Acad. Sci. USA, 76, pp. 5217-5221
- Frye, C.A., Patrick Jr., C.W., Isolation and culture of rat microvascular endothelial cells (2002) In Vitro Cell Dev. Biol., 38 A (4), pp. 208-212
- Garlanda, C., Dejana, E., Heterogeneity of endothelial cells. Specific markers (1997) Arterioscler. Thromb. Vasc. Biol., 17, pp. 1193-1202
- Garlanda, C., Parravicini, C., Sironi, M., Progressive growth in immunodeficient mice and host cell recruitment by mouse endothelial cells transformed by polyoma middle-sized T antigen: Implications of the pathogenesis of opportunistic vascular tumors (1994) Proc. Natl. Acad. Sci. USA, 91, pp. 7291-7295
- Geneser, F., Aparato circulatorio (2000) Histología. Sobre Bases Moleculares (3rd Edition), pp. 377-389. , chapter 15, ed Editorial Médica Panamericana, Buenos Aires, Argentina
- Gumkowski, F.G., Kamiska, G., Kaminski, M., Morissey, L.W., Auerbach, R., Heterogeneity of mouse vascular endothelium. In vitro studies using lymphatic, large vessels and microvascular endothelial cells (1985) Blood Vessels, 2, pp. 11-23
- Holthofer, H., Meittiner, A., Lehto, V.P., Lehtonen, E., Virtanen, I., Expression of vimentin and cytokeratin types of intermediate filament proteins in developing and adult human kidneys (1984) Lab. Invest., 50 (5), pp. 552-559
- Hughes, A.K., Ergonal, Z., Stricklett, P.K., Kohan, D.E., Molecular basis for high cells sensitivity to the cytotoxic effect of Shiga toxin-1: Upregulation of globotriaosylceramide expression (2002) J. Am. Soc. Nephrol., 13 (9), pp. 2239-2245
- Karmali, M.A., Infection by Shiga toxin-producing Escherichia coli: An overview (2004) Mol. Biotechnol., 26 (2), pp. 117-122
- Kaye, S., Louise, C., Boyd, B., Lingwood, C., Obrig, T., Shiga toxin-associated hemolytic uremic syndrome: Interleukin-1β enhancement of Shiga toxin cytotoxicity towards human vascular endothelial cells in vitro (1993) Infect. Immun., 61 (3), pp. 3886-3891
- Keusch, G.T., Acheson, D.W., Aaldering, L., Erban, J., Jacewicz, M.S., Comparison of the Shiga-like toxin 1 on cytokine- and butyrate-treated human umbilical and saphenous vein endothelial cells (1996) J. Infect. Dis., 173 (5), pp. 1164-1170
- Lanari, C., Luthy, I., Lamb, C.A., Five novel hormone-responsive cell lines derived from murine mammary ductal carcinomas: In vivo and in vitro effects of estrogens and progestins (2001) Cancer Res., 61 (1), pp. 293-302
- Langley, R., Ramirez, K., Tsan, R., Van Arsdall, M., Nilsson, M., Fidler, I., Tissue-specific microvascular endothelial cell lines from H-2K b-tsA58 mice for studies of angiogenesis and metastasis (2003) Cancer Res., 63, pp. 2971-2976
- Lidington, E.A., Moyes, D.L., McCormack, A.M., Rose, M.L., A comparison of primary endothelial cells and endothelial cell lines for studies of immune interactions (1999) Transpl. Immunol., 7 (4), pp. 239-246
- Manconi, F., Markham, R., Fraser, I., Culturing endothelial cells of microvascular origin (2000) Methods Cell Sci., 22, pp. 89-99
- Mantovani, A., Bussolino, F., Introna, M., Cytokine regulation of endothelial cell function: From molecular level to the bedside (1997) Immunol. Today, 18 (5), pp. 231-240
- Marelli-Berg, F.M., Peek, E., Lidington, E.A., Stauss, H.J., Lechler, R.I., Isolation of endothelial cells from murine tissue (2000) J. Immunol. Methods, 244 (1-2), pp. 205-215
- Milici, A.J., Furie, M.B., Carley, W.W., The formation of fenestration and channels by capillary endothelium in vitro (1985) Proc. Natl. Acad. Sci. USA, 82, pp. 6181-6185
- Morigi, M., Galbusera, M., Imberti, B., Gastoldi, S., Remuzzi, A., Zoja, A., Remuzzi, G., Verotoxin-1-induced up-regulation of adhesive molecules renders microvascular endothelial cells thrombogenic at high shear stress (2001) Blood, 98 (6), pp. 1828-1835
- Morigi, M., Micheletti, G., Figliuzzi, M., Imberti, B., Karmali, M., Remuzzi, A., Remuzzi, G., Zoja, C., Verotoxin-1 promotes leukocyte adhesion to cultured endothelial cells under physiologic flow conditions (1995) Blood, 86 (12), pp. 4553-4558
- Murray, A.B., Weibel Pallade bodies are not reliable ultrastructural markers for mouse endothelial cells (1987) Lab. Anim. Sci., 37, pp. 483-485
- Naruse, K., Fujieda, M., Miyazaki, E., An immunohistochemical study of developing glomeruli in human fetal kidneys (2000) Kidney Int., 57 (5), pp. 1836-1846
- Newman, P.J., Berndt, M.C., Gorski, J., White, G.C., Lyman, S., Paddock, C., Muller, W.A., PECAM-1 (CD31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily (1990) Science, 247 (4947), pp. 1219-1222
- Obrig, T.G., Del Vecchio, P.J., Brown, J.E., Moran, T.P., Rowland, B.M., Judge, T.K., Rothman, S.W., Direct cytotoxic action of Shiga toxin on human vascular endothelial cells (1988) Infect. Immun., 56 (9), pp. 2373-2378
- Obrig, T.G., Louise, C.B., Lingwood, C.A., Boyd, B., Barley-Maloney, L., Daniel, T.O., Endothelial heterogeneity in Shiga toxin receptors and responses (1993) J. Biol. Chem., 268 (21), pp. 15484-15488
- Ohmi, K., Kiyokawa, N., Takeda, T., Fujimoto, J., Human microvascular endothelial cells are strongly sensitive to Shiga toxins (1998) Biochem. Biophys. Res. Commun., 251, pp. 137-141
- Pijpers, A.H., Van Setten, P.A., Van Den Heuvel, L.P., Assmann, K.J., Dijkman, H.B., Pennings, A.H., Monnens, L.A., Van Hinsbergh, V.W., Verocytotoxin-induced apoptosis of human microvascular endothelial cells (2001) J. Am. Soc. Nephrol., 12 (4), pp. 767-778
- Pober, J.S., Gimbone, M.A., Lapierre, L.A., Overlapping patterns of human endothelial cells by IL-1, tumor necrosis factor and murine interferon (1986) J. Immunol., 137, pp. 1983-1986
- Porter, G.A., Palade, G.E., Milici, A.J., Differential binding of the lectin Griffonia simplicifolia and Lycopersicon esculevitum to microvascular endothelium: Organ-specific localization and partial glycoprotein characterization (1990) Eur. J. Cell Biol., 51, pp. 85-95
- Ramegowda, B., Samuel, J.E., Tesh, V.L., Interaction of Shiga toxins with human brain microvascular endothelial cells: Cytokines as sensitizing agents (1999) J. Infect. Dis., 180 (4), pp. 1205-1213
- Shappell, S.B., Mendoza, L.H., Gurpinar, T., Smith, C.W., Suki, W.N., Truong, L.D., Expression of adhesion molecules in kidney with experimental chronic obstructive uropathy: The pathogenic role of ICAM-1 and VCAM-1 (2000) Nephron, 85 (2), pp. 156-166
- Springhorn, J.P., Madri, J.A., Squinto, S.P., Human capillary endothelial cells from abdominal wall adipose tissue: Isolation using an anti-PECAM antibody (1995) In Vitro Cell Dev. Biol., 31 A (6), pp. 473-481
- Te Loo, D.M., Monnes, L.A., Van Der Velden, T.J., Vermeer, M.A., Preyers, F., Demacker, P.N., Van Den Heuvel, L.P., Van Hinsbergh, V.W., Binding and transfer of verocytotoxin by polymorphonuclear leukocytes in hemolytic uremic syndrome (2000) Blood, 95 (11), pp. 3396-3402
- Tesh, V.L., Samuel, J.E., Perera, L.P., Sharefkin, J.B., O'Brien, A., Evaluation of the role of Shiga and Shiga-like toxins in mediating direct damage to human vascular endothelial cells (1991) J. Infect. Dis., 164, pp. 344-352
- Thorin, E., Shreeve, S.M., Heterogeneity of vascular endothelial cells in normal and disease states (1998) Pharmacol. Ther., 78, pp. 155-166
- Van Setten, P.A., Van Hinsbergh, V.W., Van Der Velden, T.J., Effects of TNF alpha on verocytotoxin cytotoxicity in purified human glomerular microvascular endothelial cells (1997) Kidney Int., 51 (4), pp. 1245-1256
- Vecchi, A., Garlanda, C., Lampugnani, M.G., Monoclonal antibodies specific for endothelial cells of mouse blood vessels. Their application in the identification of adult and embryonic endothelium (1994) Eur. J. Cell Biol., 63 (2), pp. 247-254
- Voyta, J.C., Via, D.P., Butterfield, C.E., Zetter, B.R., Identification and isolation of ECs based on their increased uptake of acetylated-low-density-lipoproteins (1980) J. Cell Biol., 99, pp. 2034-2040
- Williams, J.M., Boyd, B., Nutikka, A., Lingwood, C., Barnett Foster, D., Milford, D., Mark Taylor, C., A comparison of the effects of verocytotoxin-1 on primary human renal cell cultures (1999) Toxicol. Lett., 105, pp. 47-57
- Yoshida, T., Fukuda, M., Koide, N., Ikeda, H., Sugiyama, T., Kato, Y., Ishikawa, N., Yokochi, T., Primary cultures of human endothelial cells are susceptible to low doses of Shiga toxins and undergo apoptosis (1999) J. Infect. Dis., 180, pp. 2048-2052
- Zoja, C., Angioletti, S., Donadelli, R., Shiga toxin-2 triggers endothelial leukocyte adhesion and transmigration via NF-kappaB dependent upregulation of IL-8 and MCP-1 (2002) Kidney Int., 62 (3), pp. 846-856
Citas:
---------- APA ----------
Gazzaniga, S., González, L., Mantovani, A., Vecchi, A. & Wainstok, R.
(2004)
. Isolation and molecular characterization of a mouse renal microvascular endothelial cell line. In Vitro Cellular and Developmental Biology - Animal, 40(3-4), 82-88.
http://dx.doi.org/10.1290/1543-706X(2004)040<0082:IAMCOA>2.0.CO;2---------- CHICAGO ----------
Gazzaniga, S., González, L., Mantovani, A., Vecchi, A., Wainstok, R.
"Isolation and molecular characterization of a mouse renal microvascular endothelial cell line"
. In Vitro Cellular and Developmental Biology - Animal 40, no. 3-4
(2004) : 82-88.
http://dx.doi.org/10.1290/1543-706X(2004)040<0082:IAMCOA>2.0.CO;2---------- MLA ----------
Gazzaniga, S., González, L., Mantovani, A., Vecchi, A., Wainstok, R.
"Isolation and molecular characterization of a mouse renal microvascular endothelial cell line"
. In Vitro Cellular and Developmental Biology - Animal, vol. 40, no. 3-4, 2004, pp. 82-88.
http://dx.doi.org/10.1290/1543-706X(2004)040<0082:IAMCOA>2.0.CO;2---------- VANCOUVER ----------
Gazzaniga, S., González, L., Mantovani, A., Vecchi, A., Wainstok, R. Isolation and molecular characterization of a mouse renal microvascular endothelial cell line. In Vitro Cell. Dev. Biol. Anim. 2004;40(3-4):82-88.
http://dx.doi.org/10.1290/1543-706X(2004)040<0082:IAMCOA>2.0.CO;2