Culture of endothelial cells from baboon and human glomeruli

Dollie F Green, Kay H. Hwang, Una S. Ryan, Jacques J. Bourgoignie

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Whereas mesangial and epithelial cells from glomeruli are commonly grown in vitro, there has been a failure to isolate and propagate human glomerular capillary endothelial cells. This study defines the conditions for the reproducible isolation and growth of homogeneous monolayers of primate (baboon and human) glomerular capillary endothelial cells. Using selective media and growth factors, the following criteria were identified to optimize the isolation and proliferation of glomerular endothelial cells: (1) collagenase treatment of isolated glomeruli; (2) requirement for 20% serum, endothelial cell growth factor and heparin; (3) requirement of fibronectin as surface matrix; and (4) isolation from donors less than 60 years old, as premature senescence was directly proportional to the age of the human kidney donor. Under these conditions, primary cultures with an endothelial cell composition greater than 70% were reproducibly obtained. Homogeneous endothelial monolayers were developed from 20 of 23 human kidneys, and maintained for 5 to 10 passages, depending on the age of the kidney donor. Purification to homogeneity was achieved by patch cloning or by fluorescence-activated cell sorting. Glomerular capillary endothelial cells exhibited a cobblestone morphology at confluence, expressed factor VIII-related antigen, angiotensin converting enzyme activity, and endocytosed acetylated low-density lipoproteins. Electron microscopy revealed the presence of intracellular Weibel-Palade bodies and caveolae and microvillous projections on the luminal surface. Glomerular cells also stained positive for Ulex europaeus, a lectin characteristic of human endothelial cells. In addition, preliminary results indicate that human glomerular endothelial cells increase intracellular cyGMP in response to α-human 5 to 28 atrial natriuretic peptide and intracellular free calcium in response to thrombin.

Original languageEnglish
Pages (from-to)1506-1516
Number of pages11
JournalKidney International
Volume41
Issue number6
StatePublished - Jun 1 1992

Fingerprint

Papio
Endothelial Cells
Kidney
Weibel-Palade Bodies
Endothelial Growth Factors
Caveolae
Mesangial Cells
von Willebrand Factor
Peptidyl-Dipeptidase A
Endocytosis
Fibronectins
Thrombin
Primates
Heparin
Organism Cloning
Intercellular Signaling Peptides and Proteins
Electron Microscopy
Flow Cytometry
Epithelial Cells
Calcium

ASJC Scopus subject areas

  • Nephrology

Cite this

Green, D. F., Hwang, K. H., Ryan, U. S., & Bourgoignie, J. J. (1992). Culture of endothelial cells from baboon and human glomeruli. Kidney International, 41(6), 1506-1516.

Culture of endothelial cells from baboon and human glomeruli. / Green, Dollie F; Hwang, Kay H.; Ryan, Una S.; Bourgoignie, Jacques J.

In: Kidney International, Vol. 41, No. 6, 01.06.1992, p. 1506-1516.

Research output: Contribution to journalArticle

Green, DF, Hwang, KH, Ryan, US & Bourgoignie, JJ 1992, 'Culture of endothelial cells from baboon and human glomeruli', Kidney International, vol. 41, no. 6, pp. 1506-1516.
Green DF, Hwang KH, Ryan US, Bourgoignie JJ. Culture of endothelial cells from baboon and human glomeruli. Kidney International. 1992 Jun 1;41(6):1506-1516.
Green, Dollie F ; Hwang, Kay H. ; Ryan, Una S. ; Bourgoignie, Jacques J. / Culture of endothelial cells from baboon and human glomeruli. In: Kidney International. 1992 ; Vol. 41, No. 6. pp. 1506-1516.
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