HMEC-1 - ESTABLISHMENT OF AN IMMORTALIZED HUMAN MICROVASCULAR ENDOTHELIAL-CELL LINE
HMEC-1 - ESTABLISHMENT OF AN IMMORTALIZED HUMAN MICROVASCULAR ENDOTHELIAL-CELL LINE
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DOI：
10.1111/1523-1747.ep12613748
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发表时间：
1992-12-01
影响因子：
6.5
通讯作者：
LAWLEY, TJ
中科院分区：
文献类型：
--
作者：
ADES, EW;CANDAL, FJ;LAWLEY, TJ
对人微血管内皮细胞的研究一直受到限制，因为这些细胞难以在纯培养中分离，在体外生长要求方面很苛刻，并且寿命非常有限。为了克服这些困难，我们已经转染人真皮微血管内皮细胞（HMEC）与PBR-322为基础的质粒含有编码区的猿猴病毒40 A基因产物，大T抗原，并成功地在永生化。这些细胞称为CDC/EU.HMEC-1（HMEC-1），迄今已传代95次，没有显示衰老迹象，而正常微血管内皮细胞在第8-10代时经历衰老。HMEC-1在单层培养中生长时表现出典型的鹅卵石形态，表达并分泌血管性血友病因子，摄取乙酰化低密度脂蛋白，并在基质胶上培养时快速形成管。HMEC-1的生长密度比微血管内皮细胞高3至7倍，并且需要的生长培养基严格性低得多。HMEC-1将在不存在人血清的情况下生长，而微血管内皮细胞需要补充有30%人血清的培养基。这些细胞表达通常与内皮细胞相关的其他细胞表面分子，包括CD 31和CD 36以及由单克隆抗体EN 4和PAL-E鉴定的表位。它们还表达细胞粘附分子ICAM-1和CD 44，并且在用干扰素-γ刺激后表达主要组织相容性复合物II类抗原。HMEC-1在细胞粘附测定中特异性结合淋巴细胞。因此，HMEC-1是第一个永生化的人微血管内皮细胞系，其保留了正常人微血管内皮细胞的形态、表型和功能特征。
The study of human microvascular endothelial cells has been limited, because these cells are difficult to isolate in pure culture, are fastidious in their in vitro growth requirements, and have a very limited lifespan. In order to overcome these difficulties, we have transfected human dermal microvascular endothelial cells (HMEC) with a PBR-322-based plasmid containing the coding region for the simian virus 40 A gene product, large T antigen, and succeeded in immortalizing them. These cells, termed CDC/EU.HMEC-1 (HMEC-1), have been passaged 95 times to date and show no signs of senescence, whereas normal microvascular endothelial cells undergo senescence at passages 8-10. HMEC-1 exhibit typical cobblestone morphology when grown in monolayer culture, express and secrete von Willebrand's Factor, take up acteylated low-density lipoprotein, and rapidly form tubes when cultured on matrigel. HMEC-1 grow to densities three to seven times higher than microvascular endothelial cells and require much less stringent growth medium. HMEC-1 will grow in the absence of human serum, whereas microvascular endothelial cells require culture medium supplemented with 30% human serum. These cells express other cell-surface molecules typically associated with endothelial cells, including CD31 and CD36 and epitopes identified by monoclonal antibodies EN4 and PAL-E. They also express the cell adhesion molecules ICAM-1 and CD44 and following stimulation with interferon-gamma express major histocompatibility complex class II antigens. HMEC-1 specifically bind lymphocytes in cell adhesion assays. Thus HMEC-1 is the first immortalized human microvascular endothelial cell line that retains the morphologic, phenotypic, and functional characteristics of normal human microvascular endothelial cells.