A simplified method for growth of human microvascular endothelial cells results in decreased senescence and continued responsiveness to cytokines and growth factors |
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Authors: | Birgit M Kräling Joyce Bischoff |
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Institution: | (1) Surgical Research Laboratory, Children’s Hospital, USA;(2) Department of Surgery, Harvard Medical School, 300 Longwood Avenue, 02115 Boston, Massachusetts |
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Abstract: | Summary Human dermal microvascular endothelial cells are used to analyze the functions of microvascular endothelium in vitro. However, the low yield and short lifespan of these cells in culture has limited the types of analysis that could be performed.
Human microvascular endothelial cells are typically grown in media containing supplements such as dibutyryl cyclic AMP, hydrocortisone,
bovine brain extract, and antifungal agents, each of which increase the complexity of experimental design and interpretation
of results. In the present study, endothelial cells were transferred after Ulex europeus-I selection into a simplified medium consisting of Endothelial Basal Medium 131, 10% fetal bovine serum, and 2 ng/ml basic
fibroblast growth factor and analyzed over 3 mo. The human microvascular endothelial cells expressed the endothelial markers
von Willebrand factor, CD31, P-selectin, and E-selectin. In addition, the cells showed increased proliferation in the presence
of basic fibroblast growth factor (0.5 ng/ml) or vascular endothelial cell growth factor (10 ng/ml). Tumor necrosis factor-α-induced
expression of E-selectin was similar in cells at Passages 3, 6, and 12, indicating that the cells maintained responsiveness
to cytokines over several weeks. Furthermore, the endothelial cells attained a typical cobblestone morphology with increased
cellular density and also formed capillarylike tubes on Matrigel. In summary, the human dermal microvascular endothelial cells
display the expected endothelial characteristics when grown for several passages and, therefore, provide a valuable in vitro model for human microvascular endothelium. |
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Keywords: | selectins angiogenesis tumor necrosis factor cytokines |
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