PROLIFERATION AND FUNCTION IN HUMAN ENDOTHELIAL CELLS

本研究计划的目的是研究儿童的生长和功能 从不同血管中分离培养的人内皮细胞 网站。血管组织将从脑死亡的心脏跳动中获得 身体、大血管和毛细血管内皮细胞将被隔离。在首字母中 实验将优先考虑人的髂静脉内皮细胞 来自年轻健康的个体的线条。以下目标将是 追查： 1.成人血管内皮细胞的培养和鉴定 关于EC标记功能、核型和培养寿命。 2.人血管内皮细胞体外增殖将被研究为 肝素与脑源性内皮细胞的协同作用 丝裂原、纤维连接蛋白和胶原蛋白。以下方法将是 用途：a)成分的提纯和分级；b)EC的定义 对来自a)的组件的响应；c)下列组件的相互作用 无细胞条件；d)血清对效应和相互作用的影响 组件的数量。 3.一种人类EC功能，血管紧张素转换酶的表达 (ACE)在体外，将进行研究，因为它是由合作社影响的 肝素、脑源性内皮细胞有丝分裂原、 纤维连接蛋白和胶原蛋白。将使用以下方法：a) EC对培养成分的响应(根据ACE活性)的定义 目标2；b)特异性多克隆的开发 或克隆抗体来定量ACE的数量、合成和 C)克隆和未克隆培养物中ACE表达的分析 在一个继代生长周期内。 4.在实现上述目标方面取得足够进展后， 血管部位、供者年龄和性别以及体外移植的可能影响 培养人内皮细胞的功能和增殖能力 调查过了。

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.