Carley W W, Niedbala M J, Gerritsen M E
Institute for Inflammation and Autoimmunity, Miles Research Center, West Haven, Connecticut 06516.
Am J Respir Cell Mol Biol. 1992 Dec;7(6):620-30. doi: 10.1165/ajrcmb/7.6.620.
Primary cultures of peripheral lung lobes were grown in a highly supplemented medium. Human lung endothelial cells (HLE) were isolated from the mixed population by FACS. The cells proliferated rapidly and were serially cultivated for at least 16 passages. Both early and late passage cells were positive for the standard endothelial markers. Factor VIII related-antigen (Factor VIII R-Ag), angiotensin-converting enzyme, acetylated low-density lipoprotein labeled with 1,1'-dioctadecyl-1,3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI-Ac-LDL) uptake, and bound the lectin Ulex europaeus agglutinin (UEA). Prostaglandin E2 was the major cyclooxygenase product of HLE, in contrast to human umbilical vein endothelial cells (HUVE), which synthesized PGI2 in excess of PGE2. Factor VIII R-Ag exhibited a diffuse cytoplasmic as well as an extracellular fibrillar distribution in HLE, in contrast to a vesicular (Weibel-Palade body) cytoplasmic distribution in HUVE. The HUVE did demonstrate some extracellular fibrillar Factor VIII R-Ag as well. Urokinase was the predominant plasminogen activator (PA) secreted by HLE, whereas tissue PA was predominant in HUVE cultures. HLE formed tube-like structures within 2 h of plating on a Matrigel matrix whereas HUVE formed larger tube-like structures only after 1 or more days. The properties described here indicate that human lung microvessel endothelium can be isolated and continuously grown from small tissue segments and express a number of properties that differ from those of HUVE. These studies provide further support for the concept that endothelial cells from different sources can exhibit considerable heterogeneity relating to their phenotypic and biochemical properties.
外周肺叶的原代培养物在高度补充的培养基中生长。通过荧光激活细胞分选术（FACS）从混合群体中分离出人肺内皮细胞（HLE）。这些细胞迅速增殖，并连续传代培养至少16代。早期和晚期传代细胞对标准内皮标记物均呈阳性。VIII因子相关抗原（VIII因子R-Ag）、血管紧张素转换酶、用1,1'-二辛基-1,3,3,3',3'-四甲基吲哚羰花青高氯酸盐（DiI-Ac-LDL）摄取并结合凝集素荆豆凝集素（UEA）。与合成前列环素（PGI2）超过前列腺素E2（PGE2）的人脐静脉内皮细胞（HUVE）相比，前列腺素E2是HLE的主要环氧化酶产物。VIII因子R-Ag在HLE中表现为弥漫性细胞质以及细胞外纤维状分布，而在HUVE中为囊泡状（Weibel-Palade小体）细胞质分布。HUVE也确实表现出一些细胞外纤维状VIII因子R-Ag。尿激酶是HLE分泌的主要纤溶酶原激活剂（PA），而组织型PA在HUVE培养物中占主导。HLE在铺板于基质胶基质上2小时内形成管状结构，而HUVE仅在1天或更长时间后形成更大的管状结构。这里描述的特性表明，人肺微血管内皮细胞可以从小组织片段中分离并持续生长，并表达一些与HUVE不同的特性。这些研究为不同来源的内皮细胞在表型和生化特性方面可能表现出相当大的异质性这一概念提供了进一步支持。
