Heterogeneity of microvascular endothelial cells isolated from human term placenta and macrovascular umbilical vein endothelial cells
Heterogeneity of microvascular endothelial cells isolated from human term placenta and macrovascular umbilical vein endothelial cells
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DOI：
10.1078/0171-9335-00306
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发表时间：
2003-04-01
影响因子：
6.6
通讯作者：
Desoye, G
中科院分区：
文献类型：
--
作者：
Lang, I;Pabst, MA;Desoye, G
本研究比较了人体同一器官内微血管内皮细胞和大血管内皮细胞的一些表型和生理特征。为此，采用新方法分离人足月胎盘（PLEC）微血管内皮细胞，并与人脐静脉大血管内皮细胞（HUVEC）和sv40转化的胎盘静脉内皮细胞系（HPEC-A2）进行比较。经胎盘小血管酶灌注分离PLEC，密度梯度纯化后培养。酶处理血管的组织学切片显示，灌注胎盘子叶的内皮层被选择性去除。通过内皮标志物抗血管性血血病因子、欧paeus凝集素和抗qbend10染色证实细胞的内皮身份。细胞内化乙酰化低密度脂蛋白，对巨噬细胞、平滑肌细胞和成纤维细胞的标记物没有免疫反应性。梭形PLEC以类似于静脉胎盘内皮细胞的旋涡模式生长。然而，扫描电镜检查清楚地显示PLEC在融合状态下仍然拉长，与并行研究的HPEC-A2和HUVEC的更多角形表型相反。血管活性物质（内皮素-1、2、血栓素、血管紧张素- 11、前列环素）释放到培养基中的数量和细胞因子的增殖反应与非胎儿组织衍生的人真皮微血管（MIEC）更相似，而与HUVEC更相似。有效的丝裂原如血管内皮生长因子（VEGF(121)， VEGF,65）和碱性成纤维细胞生长因子（FGF-2）诱导所有内皮细胞类型的增殖。与HUVEC（9 +/- 8%和15 +/- 20%）和HPEC-A2（15 +/- 7%和24 +/- 6%）相比，在无血清条件下培养48小时后，胎盘生长因子PIGF-1和PIGF-2能有效地刺激PLEC（142 +/- 7%和173 +/- 10%）和MIEC（160 +/- 20%和143 +/- 28%）细胞的增殖。这些数据支持以下证据：(1)本研究中描述的分离PLEC的微血管特性；(2)一个人体器官内微血管内皮细胞和大血管内皮细胞的表型和生理异质性。
The present study compares some phenotypic and physiologic characteristics of microvascular and macrovascular endothelial cells from within one human organ. To this end microvascular endothelial cells from human full-term placenta (PLEC) were isolated using a new method and compared with macrovascular human umbilical vein endothelial cells (HUVEC) and an SV40-transformed placental venous endothelial cell line (HPEC-A2). PLEC were isolated by enzymatic perfusion of small placental vessels, purified on a density gradient and cultured subsequently. Histological sections of the enzyme-treated vessels showed a selective removal of the endothelial lining in the perfused placental cotyledons. The endothelial identity of the cells was confirmed by staining with the endothelial markers anti-von Willebrand factor, Ulex europaeus lectin and anti-QBEND10. The cells internalized acetylated low-density lipoprotein and did not show immunoreactivity with markers for macrophages, smooth muscle cells and fibroblasts. The spindle-shaped PLEC grew in swirling patterns similar to that described for venous placental endothelial cells. However, scanning electron microscopic examination clearly showed that PLEC remained elongated at the confluent state, in contrast to the more polygonal phenotype of HPEC-A2 and HUVEC that were studied in parallel. The amount of vasoactive substances (endothelin-1,2, thromboxane, angiotensin 11, prostacyclin) released into the culture medium and the proliferative response to cytokines was more similar to human dermal microvessels (MIEC) derived from non-fetal tissue than to HUVEC. Potent mitogens such as vascular endothelial growth factors (VEGF(121), VEGF,65) and basic fibroblast growth factor (FGF-2) induced proliferation of all endothelial cell types. Placental growth factors PIGF-1 and PIGF-2 effectively stimulated cell proliferation on PLEC (142 +/- 7% and 173 +/- 10%) and MIEC (160 +/- 20% and 143 +/- 28%) in contrast to HUVEC (9 +/- 8% and 15 +/- 20%) and HPEC-A2 (15 +/- 7% and 24 +/- 6%) after 48h incubation time under serum-free conditions. These data support evidence for (1) the microvascular identity of the isolated PLEC described in this study, and (2) the phenotypic and physiologic heterogeneity of micro- and macrovascular endothelial cells within one human organ.