Isolation, culture, and characterization of endothelial cells from mouse glomeruli

Nese Akis, Michael P. Madaio

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Background. Cloned glomerular endothelial cells (GENC) have many potential uses and applications in immunologic and physiologic studies. Propagation of GENC has been difficult and available homogenous GENC, particularly from mice, are limited. Herein we report isolation, cloning, propagation, and characterization of GENC from mice. Methods. tsA58 immorto mice were used to isolate glomerular cells. Glomeruli were isolated by differential sieving, and decapsulated explants were cultured in permissive and optimal conditions for endothelial cells. The primary cells from glomerular outgrowths were expanded, taking advantage of the temperature-sensitive tsA58 gene, and then the cells were allowed to undergo spontaneous transformation. The cells were then sorted using anti-CD31 antibodies and their capacity to uptake acetylated-low-density lipoprotein (LDL). Individual subclones isolated by patch cloning were characterized using multiple markers. Results. One of the homogenous clones was morphologically endothelial-like, positive for CD31, CD106, CD62E, CD54, and acetylated-LDL uptake, formed tubes, and was negative for epithelial and mesangial cell markers. The functional properties of this GENC clone appeared to be intact, and signaling pathway was not altered. Two of the clones displayed the characteristics of either visceral epithelial or mesangial cells. Conclusion. The identified clones should have utility in multiple areas of investigation.

Original languageEnglish (US)
Pages (from-to)2223-2227
Number of pages5
JournalKidney International
Volume65
Issue number6
DOIs
StatePublished - Jan 1 2004

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Endothelial Cells
Clone Cells
Mesangial Cells
Organism Cloning
Epithelial Cells
Anti-Idiotypic Antibodies
Temperature
Genes
acetyl-LDL

Keywords

  • Endothelial cell
  • Glomerular cells
  • tsA58

ASJC Scopus subject areas

  • Nephrology

Cite this

Isolation, culture, and characterization of endothelial cells from mouse glomeruli. / Akis, Nese; Madaio, Michael P.

In: Kidney International, Vol. 65, No. 6, 01.01.2004, p. 2223-2227.

Research output: Contribution to journalArticle

Akis, Nese ; Madaio, Michael P. / Isolation, culture, and characterization of endothelial cells from mouse glomeruli. In: Kidney International. 2004 ; Vol. 65, No. 6. pp. 2223-2227.
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