Identification of a subpopulation of human renal microvascular endothelial cells with capacity to form capillary-like cord and tube structures |
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Authors: | Murphy Martin Harald Schoecklmann Gary Foster Lise Barley-Maloney James McKanna Thomas O Daniel |
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Institution: | (1) Department of Medicine, Vanderbilt University School of Medicine, 37232 Nashville, Tennessee;(2) Department of Medicine Cell Biology, Vanderbilt University School of Medicine, 37232 Nashville, Tennessee;(3) Medizinische Klinik IV der Universitat Erlangen-Nurnberg, Germany;(4) Nephrology Division, MCN S 3223, Vanderbilt University, 37232 Nashville, Tennessee |
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Abstract: | Summary Endothelial specialization is a prominent feature within distinct capillary beds of organs such as mammalian kidney, yet immunological
markers for functionally distinct subpopulations of cultured endothelial cells from tissue sources such as kidney have not
been available. We developed a simple and reproducible isolation and culture procedure to recover human renal microvascular
endothelial cells (HRMEC) from the cortex of unused donor kidneys. This procedure yields highly purified preparations of cells
that display endothelial markers that include Factor VIII antigen, acetyl-LDL receptors, and determinants that bind Ulex europaeus lectin. HRMEC assemble into capillary-like cord and tube structures when plated on the surface of basement membrane-like
matrix (BMM) in media containing phorbol myristate acetate.
To further define subpopulations of HRMEC, we generated a panel of monoclonal antibodies and screened for those recognizing
cell surface determinants. One monoclonal antibody recovered from this screen recognized a cell surface protein expressed
on a subpopulation of HRMEC that we have designated PEC-1 (pioneer endothelial cell antigen-1). Cells expressing PEC-1 extended
long, interconnecting filopodial processes in response to phorbol myristate acetate and assembled into capillary-like structures
when plated on BMM. Anti-PEC-1 immunoprecipitated proteins of 25 and 27 kDa. Magnetic bead separation of PEC-1 (+) cells selected
cells that assemble into capillary-like cord and tube structures. The remaining PEC-1 (−) HRMEC population formed matrix adherent
patches. In the kidney, the PEC-1 determinant is expressed on a small subpopulation of microvascular glomerular cells and
is prominently expressed on the apical membrane of proximal tubule cells. The PEC-1 determinant discriminates among subpopulations
of HRMEC, identifying a subpopulation that contributes to assembly of capillary-like structures. |
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Keywords: | endothelial heterogeneity capillary assembly marker antigen |
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