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

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。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。目前尚不存在适合重度肺动脉高压的动物模型，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影响，我们必须使用原代人类内皮细胞系。脐带静脉衬里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。我们分离这些细胞的方法是：用缓冲的生理盐水冲洗静脉，加入胰酶孵育30分钟，然后用额外的缓冲盐水冲洗/剥离细胞，然后收集细胞并将其转移到培养基中。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。从脐带中培养内皮细胞并研究其对剪切力和生长因子的反应将达到研究目标。

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.