Original Article
Copyright ©2010 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Stem Cells. Oct 26, 2010; 2(5): 114-120
Published online Oct 26, 2010. doi: 10.4252/wjsc.v2.i5.114
Spontaneous immortalization of human dermal microvascular endothelial cells
Ming Jiang, Yongfen Min, Laura DeBusk, Suzanne Fernandez, Douglas W Strand, Simon W Hayward, P Charles Lin
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
Yongfen Min, Laura DeBusk, Department of Radiation Oncology, Vanderbilt University Medical Center, 1161 21st Avenue South, Nashville, TN 37232, United States
P Charles Lin, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, United States
Author contributions: Jiang M, Min Y, DeBusk L, Fernandez S and Strand DW performed the majority of experiments and analyzed data; Lin PC provided financial support for this work; Jiang M, Hayward SW and Lin PC designed the study, analyzed data and wrote the manuscript.
Supported by (in part) Grants from NIH (CA108856, NS45888 and AR053718) to Lin PC and training grants from NIH to DeBusk L (T32CA009592)
Correspondence to: Dr. P Charles Lin, Center for Cancer Research, National Cancer Institute, Building 560, Room 12-89, Frederick, MD 21702-1201, United States. p.lin@nih.gov
Telephone: +1-301-8464688 Fax: +1-301-8467017
Received: May 20, 2010
Revised: September 22, 2010
Accepted: September 29, 2010
Published online: October 26, 2010
Abstract

AIM: To establish and characterize a spontaneously immortalized human dermal microvascular endothelial cell line, iHDME1.

METHODS: 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).

RESULTS: 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.

CONCLUSION: iHDME1 cells will be a valuable source for studying angiogenesis.

Keywords: Spontaneous immortalization; Angiogenesis; Endothelial cell