PROLIFERATION AND FUNCTION IN HUMAN ENDOTHELIAL CELLS

这项研究计划的目的是研究生长和功能 从不同血管中分离的培养的人内皮细胞（EC） 网站. 血管组织将从脑死亡的心脏跳动的 尸体和大血管和毛细血管EC将被分离。 初始 实验将优先考虑人髂静脉内皮细胞 来自年轻健康个体的品系。 以下目标将是 追求： 1.将培养成人血管EC的培养物并进行表征 关于EC标记功能、核型和培养寿命。 2.将研究人EC体外增殖， 肝素、脑源性内皮细胞 丝裂原、纤连蛋白和胶原蛋白。 将采取以下方法 使用：a）组分的纯化和分馏; B）EC的定义 对a）中各组成部分的反应; c）各组成部分在 d）血清对作用和相互作用的影响 的组件。 3.人EC功能，血管紧张素I转换酶的表达 (ACE)在体外，将进行研究，因为它是受合作 肝素，脑源性内皮细胞有丝分裂原， 纤连蛋白和胶原蛋白。 将采用以下方法： EC对培养物组分反应的定义（根据ACE活性） 在目标#2中纯化和分级; B）特异性多克隆抗体的开发 或单克隆抗体来定量ACE量，合成和 c）克隆和未克隆培养物中ACE表达的分析 在一个单一的继代培养生长周期内。 4.在实现上述目标方面取得足够进展之后， 血管部位、供体年龄和性别以及体外 培养人EC的功能和增殖， 研究了

The objective of this research program is to study the growth and function of cultured human endothelial cells (EC) isolated from diverse vascular sites. Vascular tissue will be obtained from brain-dead, heart-beating cadavers, and large vessel and capillary EC will be isolated. In initial experiments priority will be given to a human iliac vein endothelial cell line derived from a young, healthy individual. The following goals will be pursued: 1. Cultures of adult human vascular EC will be cultivated and characterized with respect to EC marker functions, karyotypes, and culture lifespans. 2. Human EC proliferation in vitro will be investigated as effected by cooperative interactions between heparin, brain-derived endothelial mitogen, fibronectin, and collagen. The following approaches will be used: a) Purification and fractionation of components; b) Definition of EC response to components from a); c) Interaction of components under cell-free conditions; d) Influence of serum on the effects and interaction of components. 3. A human EC function, the expression of angiotensin I-converting enzyme (ACE) in vitro, will be investigated as it is affected by cooperative interactions between heparin, brain-derived endothelial mitogen, fibronectin, and collagen. The following approaches will be used: a) Definition of EC response (in terms of ACE activity) to culture components purified and fractionated in Aim #2; b) Development of specific polyclonal or monoclonal antibodies to quantitate ACE amounts, synthesis and degradation; c) Analyses of ACE expression in cloned and uncloned cultures within a single subcultivation growth cycle. 4. After sufficient progress has been made in achieving the above goals, possible effects of vascular site, donor age and sex, and in vitro cultivation on function and proliferation in culture human EC will be investigated.