Isolation and characterization of CD133+CD34+VEGFR-2+CD45− fetal endothelial cells from human term placenta

• Published in: Microvascular research Vol. 84; no. 1; pp. 65 - 73
• Main Authors: Sölder, Elisabeth, Böckle, Barbara C., Nguyen, Van Anh, Fürhapter, Christina, Obexer, Petra, Erdel, Martin, Stössel, Hella, Romani, Nikolaus, Sepp, Norbert T.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-07-2012
• Subjects: AC133 Antigen; Adult; Antigens, CD - metabolism; Antigens, CD34 - metabolism; Biomarkers - metabolism; Cell Culture Techniques; Cells, Cultured; Cytoplasmic Structures - ultrastructure; Dermis - blood supply; Endothelial Cells - cytology; Endothelial Cells - metabolism; Endothelial Cells - ultrastructure; Female; Glycoproteins - metabolism; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Leukocyte Common Antigens - metabolism; Microvilli - ultrastructure; Peptides - metabolism; Placenta - blood supply; Pregnancy; Term Birth; Vascular Endothelial Growth Factor Receptor-2 - metabolism
• ISSN: 0026-2862; 1095-9319
• DOI: 10.1016/j.mvr.2012.03.005

The phenotypes and functions of endothelial cells (EC), a heterogeneous cell population, vary along the vascular tree and even in the same organ between different vessels. The placenta is an organ with abundant vessels. To enhance further knowledge concerning placenta derived EC, we develop a new method for isolation, purification and culture of these EC. Moreover, in order to investigate the peculiarity of placenta derived EC we compare their phenotypic and functional characteristics with human dermal lymphatic endothelial cells (HDLEC) and human umbilical vein endothelial cells (HUVEC). Freshly isolated placenta derived EC displayed an elongated shape with pale cytoplasm and showed the typical cobblestone pattern of EC but also a swirling pattern when confluent. FISH-analyses of the isolated EC from placentae of male fetus revealed an XY genotype strongly indicating their fetal origin. Characterisation of placenta derived fetal EC (fEC) underlined their blood vessel phenotype by the expression of vWF, Ulex europaeus lectin-1, HLA-class I molecules, CD31, CD34, CD36, CD51/61, CD54, CD62E, CD105, CD106, CD133, CD141, CD143, CD144, CD146, VEGFR-1, VEGFR-2, EN-4, PAL-E, BMA120, Tie-1, Tie-2 and α-Tubulin. In contrast to previous reports the expression of lymphatic markers, like VEGFR-3, LYVE-1, Prox-1 and Podoplanin was consistently negative. Haematopoietic surface markers like CD45 and CD14 were also always negative. Various functional tests (Dil-Ac-LDL uptake, Matrigel assay and TNF-α induced upregulation of CD62E and CD54) substantiated the endothelial nature of propagated fEC. At the ultrastructural level, fEC harboured numerous microvilli, micropinocytic vesicles at their basis, were rich in intermediate filaments and possessed typical Weibel - Palade bodies. In conclusion, the placenta is a plentiful source of fetal, microvascular, blood EC with an expression profile (CD34+, CD133+, VEGFR-2+, CD45-) suggestive of an endothelial progenitor phenotype.
► Phenotype and function of endothelial cells vary along the vascular tree. ► Efficient method for human placental endothelial cell propagation. ► Characterisation of placental microvascular endothelial cells and comparison with other endothelial cell lines. ► Blood but not lymphatic endothelial marker expression found in placental microvascular endothelial cells. ► Placental microvascular endothelial cells express progenitor cell markers, but not haematopoietic surface markers.