HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell Line

Ades, Edwin W.; Candal, Francisco J.; Swerlick, Robert A.; George, Velma G.; Summers, Susan; Bosse, Diane; Lawley, Thomas J.

doi:10.1111/1523-1747.ep12613748

Cited by 1,160 publications

(868 citation statements)

References 41 publications

Supporting

Mentioning

852

Contrasting

Unclassified

Order By: Relevance

“…HMEC-1 (SV40 T-antigen transformed human dermal microvascular endothelial cell) 27 and HUVEC (ATCC CRL-1730) were maintained in microvascular endothelial cell growth medium (Clonetics Corporation, obtained from TCS Biologicals, Buckingham, UK).…”

Section: Methodsmentioning

confidence: 99%

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

2000

View full text Add to dashboard Cite

Tumour growth is dependent upon a blood supply and is associated with the switch to the angiogenic phenotype. We are developing strategies for targeting gene expression to endothelial cells in the tumour vasculature. Recombinant retroviruses have been generated that incorporate regulatory sequences of the prepro-endothelin-1 (ppET1) promoter. Following reverse transcription and integration these modifications are duplicated in the proviral 5Ј LTR for transcription of the internal ␤-galactosidase reporter gene. The titres and endothelial specificity of retroviral vectors harbouring different modifications have been analysed. In the optimal strategy, replacing the MLV enhancer with ppET1 promoter sequences containing the GATA and AP1 elements whilst

show abstract

Section: Methodsmentioning

confidence: 99%

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

2000

View full text Add to dashboard Cite

show abstract

“…The human microvascular endothelial cell line HMEC-1 was kindly provided by Edwin Ades (Centers for Disease Control and Prevention, Atlanta, GA). 35,36 HMEC-1 cells were cultured in MDCB131 medium with 10% FCS. hFOB 1.19 cells (ATCC, Manassas, VA) were cultured in DMEM/F12 medium without Phenol Red with 10% FCS.…”

Section: Cells and Cell Culturementioning

confidence: 99%

Identification of Oncostatin M as a STAT5-Dependent Mediator of Bone Marrow Remodeling in KIT D816V-Positive Systemic Mastocytosis

Hoermann

Cerny-Reiterer

Perné

et al. 2011

The American Journal of Pathology

View full text Add to dashboard Cite

Systemic mastocytosis is a neoplastic disease of mast cells harboring the activating KIT mutation D816V. In most patients, mast cell infiltration in the bone marrow is accompanied by marked microenvironment alterations, including increased angiogenesis, osteosclerosis, and sometimes fibrosis. Little is known about the mast cell-derived molecules contributing to these bone marrow alterations. We show here that neoplastic mast cells in patients with systemic mastocytosis express oncostatin M (OSM), a profibrogenic and angiogenic modulator. To study the regulation of OSM expression, KIT D816V was inducibly expressed in Ba/F3 cells and was found to up-regulate OSM mRNA and protein levels, suggesting that OSM is a KIT D816V-dependent mediator. Correspondingly, KIT D816V ؉ HMC-1.

show abstract

“…Endothelial cells from human dermis immortalized with SV40 (HMEC) were originally obtained from Dr. T.J. Lawley, Biological Products Branch, Centers for Disease Control, Atlanta, Georgia [26]. Cells were maintained in endothelial basal medium MCDB131 (Gibco/ BRL, Grand Island, NY) and supplemented with 15% fetal bovine serum (FBS) (Sigma, St. Louis, MO), 10 ng/ml epidermal growth factor (Collaborative Biochemical Products, Bedford, MA), 1 μg/ml hydrocortisone (Sigma), penicillin, and streptomycin (Gibco/BRL).…”

Section: Cells and Culture Conditionsmentioning

confidence: 99%

Delayed radioprotection by nuclear transcription factor κB -mediated induction of manganese superoxide dismutase in human microvascular endothelial cells after exposure to the free radical scavenger WR1065

Murley

Kataoka

Weydert

et al. 2006

Free Radical Biology and Medicine

View full text Add to dashboard Cite

The free radical scavenger WR1065 (SH) is the active thiol form of the clinically approved cytoprotector amifostine. At doses of 40 μM and 4 mM it can activate the redox-sensitive nuclear transcription factor κB (NFκB) and elevate the expression of the antioxidant gene manganese superoxide dismutase (MnSOD) in human microvascular endothelial cells (HMEC). MnSOD contains binding motifs for a number of transcription factors other than NFκB and codes for a potent antioxidant enzyme localized in the mitochondria that is known to confer enhanced radiation resistance to cells. The purpose of this study was to determine the effect of WR1065 exposure on the various transcription factors known to affect MnSOD expression along with the subsequent kinetics of intracellular elevation of MnSOD protein levels and associated change in sensitivity to ionizing radiation in HMEC. Cells were grown to confluence and exposed to WR1065 for 30 min. Affects on the transcription factors AP1, AP2, CREB, NFκB, and Sp1 were monitored as a function of time ranging from 30 min to 4 h after drug exposure using a gel-shift assay. Only NFκB exhibited a marked activation and that occurred 30 min following the cessation of drug exposure. MnSOD protein levels, as determined by Western blot analysis, increased up to 16-fold over background control levels by 16 h following drug treatment, and remained at 10-fold or higher levels for an additional 32 h. MnSOD activity was evaluated using a gel-based assay and was found to be active throughout this time period. HMEC were irradiated with X-rays either in the presence of 40 μM or 4 mM WR1065 or 24 h after its removal when MnSOD levels were most elevated. No protection was observed for cells irradiated in the presence of 40 μM WR1065. In contrast, a 4 mM dose of WR1065 afforded an increase in cell survival by a factor of 2. A "delayed radioprotective" effect was, however, observed when cells were irradiated 24 h later, regardless of the concentration of WR1065 used. This effect is characterized as an increase in survival at the 2 Gy dose point, i.e., a 40% increase in survival, and an increase in the initial slope of the survival curve by a factor of 2. The anti-inflammatory sesquiterpene lactone, Helenalin, is an effective inhibitor of NFκB activation. HMEC were exposed to Helenalin for 2 h at a nontoxic concentration of 5 μM prior to exposure to WR1065. This treatment not only inhibited activation of NFκB by WR1065, but also inhibited the subsequent elevation of MnSOD and the delayed radioprotective effect. A persistent marked elevation of MnSOD in cells following their exposure to a thiol-containing reducing agent such as WR1065 can result in an elevated resistance to the cytotoxic effects of ionizing radiation and represents a novel radioprotection paradigm.

show abstract

HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell Line

Cited by 1,160 publications

References 41 publications

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

Identification of Oncostatin M as a STAT5-Dependent Mediator of Bone Marrow Remodeling in KIT D816V-Positive Systemic Mastocytosis

Delayed radioprotection by nuclear transcription factor κB -mediated induction of manganese superoxide dismutase in human microvascular endothelial cells after exposure to the free radical scavenger WR1065

Contact Info

Product

Resources

About