Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro.
EC culture models are essential to study pathological alterations of endothelial cells (ECs) in pulmonary vascular diseases under standardized conditions. Nevertheless, little is known about the spectrum of alterations of vessel-specific endothelial phenotypes in monolayer cultures. For the comparative study of endothelial markers in vivo and in vitro we investigated immunohistochemically the expression of PECAM-1, vWf, and CD34 by pulmonary ECs in vivo and in stimulated/unstimulated human umbilical vein endothelial cells (HU-VEC) and human pulmonary microvascular endothelial cells (HPMEC). In vivo, vessel type-specific expression patterns were found for vWf and CD34, while PECAM-1 was homogeneously and strongly expressed. While all HUVEC showed a marked vWf staining, about two-thirds of HPMEC exhibited a strong and the rest a moderate vWf staining. In both in vitro models all ECs were clearly PECAM-1-positive. However, only about 20% of the HUVEC and HPMEC were CD34-positive. Our results demonstrate the reduced expression of vessel type-specific endothelial phenotypes by endothelial monolayer cultures, stressing the need to improve culture conditions as well as develop cocultures and three-dimensional culture models. Moreover, the need for endothelial markers specific for single microvascular type ECs becomes obvious in order to establish cultures consisting of only one microvascular ECs subpopulation.
Müller
Annette M
AM
Institute of Pathology, University-Clinic Bergmannsheil, Ruhr-University Bochum, Bürkle-de-la-Camp-Platz 1, Bochum, 44789 Germany. annette.mueller@ruhr-uni-bochum.de
Hermanns
M Iris
MI
Skrzynski
Carmen
C
Nesslinger
Maya
M
Müller
Klaus-Michael
KM
Kirkpatrick
C James
CJ
D018952:Antigens, CD34 / Q000378:metabolism*; D015415:Biomarkers; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000033:anatomy & histology / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008168:Lung / Q000098:blood supply / Q000276:immunology / Q000378:metabolism; D008833:Microcirculation / Q000033:anatomy & histology / Q000276:immunology / Q000378:metabolism; D010641:Phenotype; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism*; D014841:von Willebrand Factor / Q000378:metabolism*
D018952:Antigens, CD34; D015415:Biomarkers; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014841:von Willebrand Factor