Ming Jiang, Suzanne Fernandez, Douglas W Strand, Simon W Hayward, Department of Urologic Surgery, Vanderbilt University Medical Center, 1161 21st Avenue South, Nashville, TN 37232, United States.
World J Stem Cells. 2010 Oct 26;2(5):114-20. doi: 10.4252/wjsc.v2.i5.114.
To establish and characterize a spontaneously immortalized human dermal microvascular endothelial cell line, iHDME1.
We developed a spontaneous immortalization method. This approach is based on the application of optimized culture media and culture conditions without addition of any exogenous oncogenes or carcinogens. Using this approach, we have successfully established a microvascular endothelial cell line, iHDME1, from primary human dermal microvascular endothelial cells. iHDME1 cells have been maintained in culture dishes for more than 50 passages over a period of 6 mo. Using a GFP expressing retrovirus, we generated a GFP-stable cell line (iHDME1-GFP).
iHDME1 retain endothelial morphology and uniformly express endothelial markers such as VEGF receptor 2 and VE-cadherin but not α-smooth muscle actin (α-SM-actin) and cytokeratin 18, markers for smooth muscle cells and epithelial cells respectively. These cells retain endothelial properties, migrate in response to VEGF stimulation and form 3-D vascular structures in Matrigel, similar to the parental cells. There is no significant difference in cell cycle profile between the parental cells and iHDME1 cells. Further analysis indicates enhanced stemness in iHDME1 cells compared to parental cells. iHDME1 cells display elevated expression of CD133 and hTERT.
iHDME1 cells will be a valuable source for studying angiogenesis.
建立并鉴定一个自发永生化的人真皮微血管内皮细胞系，即 iHDME1。
我们开发了一种自发永生化方法。该方法基于优化的培养体系和培养条件，不添加任何外源性癌基因或致癌物质。采用该方法，我们成功地从原代人真皮微血管内皮细胞中建立了一个微血管内皮细胞系，即 iHDME1。iHDME1 细胞在培养皿中已维持培养超过 50 代，历时 6 个月。我们使用表达 GFP 的逆转录病毒，生成了 GFP 稳定的细胞系（iHDME1-GFP）。
iHDME1 保留了内皮细胞的形态，均匀表达内皮标志物，如 VEGF 受体 2 和 VE-钙黏蛋白，但不表达平滑肌肌动蛋白（α-SM-actin）和细胞角蛋白 18，分别是平滑肌细胞和上皮细胞的标志物。这些细胞保留了内皮细胞的特性，能在 VEGF 刺激下迁移，并在 Matrigel 中形成 3-D 血管结构，与亲本细胞相似。亲本细胞和 iHDME1 细胞的细胞周期特征没有明显差异。进一步的分析表明，与亲本细胞相比，iHDME1 细胞的干性增强。iHDME1 细胞表现出 CD133 和 hTERT 的高表达。
iHDME1 细胞将成为研究血管生成的有价值的细胞来源。
