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

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 重度肺动脉高压时存在肺血管系统重构。 这种重塑涉及内皮细胞的结构和表型改变。 目前还没有合适的重度肺动脉高压动物模型，因此我们仅限于使用组织培养模型和人体组织。 为了测试不同生长因子的作用，我们必须使用原代人内皮细胞系。脐带静脉内的细胞易于分离和培养，在培养中存活良好，是典型的动脉内皮细胞。 我们通过用缓冲盐水溶液冲洗静脉，加入胰蛋白酶孵育30分钟，然后用额外的缓冲盐水冲洗/剥离细胞，然后收集并转移到培养基中来分离这些细胞。 本研究的目的是收集原代人内皮细胞，以便在体外模型中研究肺动脉高压。 培养脐带内皮细胞并研究其对切应力和生长因子的反应将达到研究目的。

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.