title: Spontaneous immortalization of human dermal microvascular endothelial cells.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  iHDME1 cells will be a valuable source for studying angiogenesis.
authors: 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.
authors: Ming
authors: M
authors: Jiang
authors: Yongfen
authors: Y
authors: Min
authors: Laura
authors: L
authors: Debusk
authors: Suzanne
authors: S
authors: Fernandez
authors: Douglas W
authors: DW
authors: Strand
authors: Simon W
authors: SW
authors: Hayward
authors: P Charles
authors: PC
authors: Lin
references: Carcinogenesis. 1998 Apr;19(4):673-81
references: 9600354
references: Mol Cell Biol. 2000 Feb;20(4):1436-47
references: 10648628
references: Cancer Res. 1999 Jun 1;59(11):2516-21
references: 10363964
references: Transplantation. 2004 May 15;77(9):1357-65
references: 15167590
references: In Vitro Cell Dev Biol Anim. 2002 Mar;38(3):165-72
references: 12026165
references: Cell. 2000 Aug 18;102(4):407-10
references: 10966103
references: Cell. 1976 Dec;9(4 PT 2):685-93
references: 189941
references: Oncogene. 1999 Dec 13;18(53):7676-80
references: 10618707
references: Science. 2001 Feb 2;291(5505):868-71
references: 11157165
references: J Cell Biol. 1988 Mar;106(3):761-71
references: 2450098
references: J Clin Invest. 2008 Jul;118(7):2592-9
references: 18535669
references: Mol Cells. 2002 Jun 30;13(3):351-61
references: 12132573
references: Stem Cells. 2010 Feb;28(2):344-56
references: 20020426
references: Science. 1998 Jan 16;279(5349):349-52
references: 9454332
references: J Invest Dermatol. 2000 Mar;114(3):444-55
references: 10692102
references: Cancer Res. 1990 Sep 15;50(18):6075-86
references: 1975513
references: Exp Cell Res. 1961 Dec;25:585-621
references: 13905658
references: J Invest Dermatol. 1992 Dec;99(6):683-90
references: 1361507