ISOLATION AND GROWTH OF UMBILICAL VEIN ENDOTHELIAL CELLS IN VITRO FOR STUDIES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There is remodeling of the pulmonary vascular system in severe pulmonary hypertension. This remodeling involves structural and phenotypic alterations of endothelial cells. There does not yet exist a suitable animal model for severe pulmonary hypertension, so we are limited to using tissue culture models and human tissue. In order to test the effects of different growth factors we must use a primary human endothelial cell line. The cells lining the vein of an umbilical cord are easy to isolate and culture, survive well in culture and are typical of arterial endothelial cells. We isolate these cells by rinsing the vein with a buffered saline solution, adding trypsin for a 30-minute incubation and then rinsing/stripping the cells out with additional buffered saline before collecting and transferring them to a culture media. The objective of this study is to collect primary human endothelial cells in order to study pulmonary hypertension in an in vitro model. Culturing endothelial cells from umbilical cords and studying their response to shear stress and growth factors will achieve the study objectives.