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Isolation Culture and Characterization of Endothelial Cells from Mouse Glomeruli       
Yazarlar (2)
Prof. Dr. Neşe SANSA Prof. Dr. Neşe SANSA
Haliç Üniversitesi, Türkiye
Michael P. Madaio
Devamını Göster
Özet
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.
Anahtar Kelimeler
Endothelial cell | Glomerular cells | tsA58
Makale Türü Açık Erişim Özgün Makale
Makale Alt Türü SSCI, AHCI, SCI, SCI-Exp dergilerinde yayınlanan tam makale
Dergi Adı Kidney International
Dergi ISSN 0085-2538 Wos Dergi Scopus Dergi
Dergi Tarandığı Indeksler SCI-Expanded
Dergi Grubu Q4
Makale Dili İngilizce
Basım Tarihi 01-2004
Cilt No 65
Sayı 6
Sayfalar 2223 / 2227
Doi Numarası 10.1111/j.1523-1755.2004.00634.x