Isolation and molecular characterization of a mouse renal microvascular endothelial cell line

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 isol...

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Autores principales:	Gazzaniga, S., González, L., Mantovani, A., Vecchi, A., Wainstok, R.
Formato:	JOUR
Materias:	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
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_10712690_v40_n3-4_p82_Gazzaniga
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_10712690_v40_n3-4_p82_Gazzaniga2023-10-03T16:02:38Z Isolation and molecular characterization of a mouse renal microvascular endothelial cell line Gazzaniga, S. González, L. Mantovani, A. Vecchi, A. Wainstok, R. 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 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. Fil:Gazzaniga, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_10712690_v40_n3-4_p82_Gazzaniga
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 Gazzaniga, S. González, L. Mantovani, A. Vecchi, A. Wainstok, R. Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
topic_facet	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
description	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.
format	JOUR
author	Gazzaniga, S. González, L. Mantovani, A. Vecchi, A. Wainstok, R.
author_facet	Gazzaniga, S. González, L. Mantovani, A. Vecchi, A. Wainstok, R.
author_sort	Gazzaniga, S.
title	Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
title_short	Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
title_full	Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
title_fullStr	Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
title_full_unstemmed	Isolation and molecular characterization of a mouse renal microvascular endothelial cell line
title_sort	isolation and molecular characterization of a mouse renal microvascular endothelial cell line
url	http://hdl.handle.net/20.500.12110/paper_10712690_v40_n3-4_p82_Gazzaniga
work_keys_str_mv	AT gazzanigas isolationandmolecularcharacterizationofamouserenalmicrovascularendothelialcellline AT gonzalezl isolationandmolecularcharacterizationofamouserenalmicrovascularendothelialcellline AT mantovania isolationandmolecularcharacterizationofamouserenalmicrovascularendothelialcellline AT vecchia isolationandmolecularcharacterizationofamouserenalmicrovascularendothelialcellline AT wainstokr isolationandmolecularcharacterizationofamouserenalmicrovascularendothelialcellline
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Isolation and molecular characterization of a mouse renal microvascular endothelial cell line

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