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 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 alpha-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.
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina.
Silvina
S
Gazzaniga
Lorena
L
González
Alberto
A
Mantovani
Annunciata
A
Vecchi
Rosa
R
Wainstok
D000199:Actins / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D007633:Keratins / Q000378:metabolism; D007668:Kidney / Q000098:blood supply*; D008070:Lipopolysaccharides / Q000633:toxicity; D008077:Lipoproteins, LDL / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012692:Serial Passage / Q000379:methods; D022622:Shiga Toxin 1 / Q000633:toxicity; D014709:Vero Cells; D014746:Vimentin / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
D000199:Actins; D008070:Lipopolysaccharides; D008077:Lipoproteins, LDL; D022622:Shiga Toxin 1; D014746:Vimentin; C032629:acetyl-LDL; D014841:von Willebrand Factor; D007633:Keratins