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

Silvina Gazzaniga, Lorena González, Alberto Mantovani, Annunciata Vecchi, Rosa Wainstok

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)82-88
Number of pages7
JournalIn Vitro Cellular & Developmental Biology - Animal
Volume40
Issue number3-4
DOIs
Publication statusPublished - Mar 2004

Keywords

  • CD31
  • Endothelium
  • ICAM
  • Shiga toxin
  • VCAM

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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