Primary Human Brain Microvascular Endothelial Cells (ACBRI 376) - Cell Systems
Primary Human Brain Microvascular Endothelial Cells (ACBRI 376) were initiated by elutriation of dispase-dissociated normal human brain cortex tissue.
Primary Human Brain Microvascular Endothelial Cells can provide a useful tool for adherence, transport and permeability studies of the blood-brain-barrier (BBB). ACBRI 376 cells demonstrate particular markers of differentiation (interdigitated cell contact, desmosomes, Z0-1 protein epitopes) similar to those observed in vivo. The morphological and immunofluorescent markers detailed for ACBRI 376 are suggestive of a differentiated cell monolayer that can be used to study properties of the human BBB.
ACBRI 376 cells 3 days after plating, plated with Attachment Factor™, cultured in Cell Systems Complete Medium Kit With Serum and CultureBoost-R™ (4Z0-500-R).
ACBRI 376 cells express endothelial cell markers including CD31 and von Willebrand Factor. They also take up DiI-Ac-LDL in an endothelial cell functional assay.
ACBRI 376 cells express ZO-1 indicating the formation of tight junctions.
These cells were originated using Cell Systems Complete Serum-Free Medium (SF-4Z0-500), and subsequently grown and passaged in Cell Systems Complete Medium (4Z0-500). They are available at Passage 3 [< 12 cumulative population doublings] cryopreserved in Cell Systems Cell Freezing Medium™ (4Z0-705). This vial will initiate a Passage 4 cell culture in a 75cm2 flask.
In addition to cryopreserved vials, these cells also are available in 25cm2 and 75cm2 proliferating cell culture flasks (US domestic market only).
Each vial of cells is shipped with vials of Bac-Off® (antibiotic) and CultureBoost™ (animal derived growth factors) or CultureBoost-R™ (human recombinant growth factors) at no additional cost.
These cells are qualified for use with:
HIV Serologic Test (donor level HIV AB EIA)
HIV PCR Test (frozen cell pool by CLIA Licensed Clinical Lab)
Hepatitis B (HBV) and Hepatitis C (HCV) PCR Test (at frozen cell level)
|Test of frozen cells for Mycoplasma spp. (ATCC method by CLIA Licensed Clinical Lab)||Negative|
Cell Markers and Functional Tests
|CD31||> 98% positive by immunofluorescence|
VWF / Factor VIII
> 95% positive by immunofluorescence
Uptake of Di-I-Ac-LDL
> 95% positive by immunofluorescence
Characterization in Peer-Reviewed Literature
Modified with permission from Storm et al EMBO Molecular Medicine (2019) 11:e9164.
Figure S1. Characterisation of HBMEC (Cell Systems ACBRI 376) at number of passages, as measured by flow cytometry. Unlabelled cells are depicted in grey. Blue = passage 5, light green = passage 7 (only in B), red = passage 8, dark green = passage 9. A, Uptake of DiI-labelled acetylated low density lipoprotein. B, Levels of CD31 expression. C, Upregulation of ICAM-1 by 10ng/ml TNF; dotted lines represent unstimulated labelled cells and the continuous lines represent the TNF-stimulated cells. D, Levels of EPCR expression on TNF-stimulated HBMEC. E, Levels of CD36 expression on TNF-stimulated HBMEC.
Reprinted with permission from Smits and Mir et al. PLoS ONE (2011) 6:e16282. Figure 5. Inhibition of EZH2 reduces endothelial tubule formation in vitro. (A) HBMVECs (ACBRI 376) were cultured on Matrigel coated plates in EBM, EGM, or EBM supplemented with U87-GFP cells. Scale bar = 450 µm. Inhibition of EZH2 in HBMVECs, either by transfection with pre-miR-101, EZH2 siRNA, or treatment with DZNep significantly reduced tubule formation as compared to control cells. Tubule formation was assessed as tubule length and branching. (B) Quantitation of tubule length and branching using ImageJ software. (n = 3) Error bars indicate s.d. *p<0.05, ***p<0.001, t test.
Reprinted with permission from Smits and Mir et al. PLoS ONE (2011) 6:e16282. Figure 6. Inhibition of EZH2 reduces endothelial migration and invasion in vitro. (A) HBMVEC (ACBRI 376) monolayer cultures were scratched. Images were acquired directly after scratching (t = 0) and 24 h later (t = 24). The migration front is indicated by the dashed lines. Scale bar = 450 µm. Inhibition of EZH2 in HBMVECs, either by transfection with pre-miR-101, EZH2 siRNA or treatment with DZNep significantly reduced migration as compared to control. (B) Quantitation of endothelial cell migration into the scratched area using ImageJ software. (C and D) HBMVECs were transfected with pre-miR-101, EZH2 siRNA, non-related control molecules, or treated with DZNep, incubated on a Transwell system and subsequently analyzed for invasion capability. EZH2 inhibition, either through pre-miR-101, EZH2 siRNA or DZNep, significantly decreased invasion as shown by Hoechst staining. Scale bar = 225 µm. (n = 3) Error bars indicate s.d. *p<0.05, ***p<0.001, t test.
Reprinted with permission from Niego et al. PLoS ONE (2017) 12: e0177332.
Figure 5. Only KD025, but not fasudil, protects brain endothelial cells (ACBRI 376) from rt-PA and plasmin. (A) Small molecule KD025 concentration-dependently blocks permeability changes in primary human brain microvascular endothelial cells (ACBRI 376 BECs; cultured without astrocytes) treated for 6 h with DMSO as control or with rt-PA (25nM) and human plasminogen (plgn; 100nM), with or without KD025 (2 and 20μM). n = 4. (B) Comparison of selective ROCK-2 inhibition by KD025 (20μM) versus non-selective ROCK inhibition by fasudil (HA1077; 2, 20 and 100μM) against rt-PA+plasminogen (25nM+100nM, respectively) in BECs, as assessed 6 h post stimulation. HA1077 displays no protective capacity in BECs. n = 3. Bars represent mean±SEM. **P<0.01 compared to all other groups, ***P<0.001 compared to t-PA+Plgn by one-way ANOVA with Tukey’s post hoc analysis. ## P<0.01 compared to DMSO control and P = 0.06 by two-tailed paired t-test. (C) Representative phase-contrast micrographs (top panels) and double immunofluorescence images of zonula occludens 1 (ZO-1, representing tight junctions (TJs); white) and nuclei (Hoechst; red) (bottom panels) of ACBRI 376 BECs stimulated for 6 h with DMSO (control) or with rt-PA+plasminogen (25nM+100nM, respectively), in the presence or absence of KD025 (20μM). KD025 attenuates morphological changes (arrows) and preserves TJs in endothelial cells. Asterisks depict areas with degraded TJs. Scale bars = 200μm.
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- "Tick-borne encephalitis virus infects human brain microvascular endothelial cells without compromising blood-brain barrier integrity" Palus et al. Virology, 2017
- "Selective inhibition of brain endothelial Rho-kinase-2 provides optimal protection of an in vitro blood-brain barrier from tissue-type plasminogen activator and plasmin" Niego and Lee et al. PLoS ONE 2017
- "α-Klotho expression determines nitric oxide synthesis in response to FGF-23 in human aortic endothelial cells" Chung et al. PLOS One, 2017
- "Tetrahydroxy stilbene glucoside ameliorates H2O2-induced human brain microvascular endothelial cell dysfunction in vitro by inhibiting oxidative stress and inflammatory responses" Jiang et al. Molecular Medicine Reports, 2017.
- "Overexpression of pregnane X and glucocorticoid receptors and the regulation of cytochrome P450 in human epileptic brain endothelial cells" Ghosh et al. Epilepsia, 2017.
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- "Long noncoding RNA MALAT1 inhibits apoptosis induced by oxygen-glucose deprivation and reoxygenation in human brain microvascular endothelial cells" Xin and Jiang, Experimental and Therapeutic Medicine, 2017.
- "VEGFR2 alteration in Alzheimer’s disease" Cho et al. Scientific Reports, 2017.
- "Tumour necrosis factor‐α‐mediated disruption of cerebrovascular endothelial barrier integrity in vitro involves the production of proinflammatory interleukin‐6" Rochfort et al. J Neurochemistry, 2016
- "Neuropilin-1 modulates interferon-γ-stimulated signaling in brain microvascular endothelial cells" Wang et al. Journal of Cell Science, 2016
- "Cerebral ischemia-induced angiogenesis is dependent on tumor necrosis factor receptor 1-mediated upregulation of α5β1 and αVβ3 integrins." Huang et al. J Neuroinflammation, 2016.
- "Shiga Toxin Mediated Neurologic Changes in Murine Model of Disease" Pradhan et al. Frontiers in Cellular and Infection Microbiology, 2016.
- "Tumor-Derived Factors and Reduced p53 Promote Endothelial Cell Centrosome Over-Duplication" Yu et al. PLoS One, 2016.
- "Different Regulation of p53 Expression by Cadmium Exposure in Kidney, Liver, Intestine, Vasculature, and Brain Astrocytes" Lee et al. Toxicology Research, 2016.
- "Endothelial progenitor cells and neural progenitor cells synergistically protect cerebral endothelial cells from Hypoxia/reoxygenation-induced injury via activating the PI3K/Akt pathway" Wang et al. Molecular Brain, 2016.
- "Distinct Contributions of Astrocytes and Pericytes to Neuroinflammation Identified in a 3D Human Blood-Brain Barrier on a Chip" Herland et al. PLoS One, 2016.
- "The Novel Methods for Analysis of Exosomes Released from Endothelial Cells and Endothelial Progenitor Cells" Wang et al. Stem Cells International, 2016.
- "Rapid endothelial cytoskeletal reorganization enables early blood–brain barrier disruption and long-term ischaemic reperfusion brain injury" Shi et al. Nature Communications, 2016.
- "Interaction between Endothelial Protein C Receptor and Intercellular Adhesion Molecule 1 to Mediate Binding of Plasmodium falciparum-Infected Erythrocytes to Endothelial Cells" Avril et al. mBio, 2016.
- "Optimization of an in vitro human blood–brain barrier model: Application to blood monocyte transmigration assays" Paradis et al. MethodsX, 2016.
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- "Retinal pericytes and cytomegalovirus infectivity: implications for HCMV-induced retinopathy and congenital ocular disease" Wilkerson et al. J Neuroinflammation, 2015
- "Brain vascular pericytes following ischemia have multipotential stem cell activity to differentiate into neural and vascular lineage cells" Nakagomi et al. Stem Cells, 2015
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"JAM-A and ALCAM are therapeutic targets to inhibit diapedesis across the BBB of CD14+CD16+ monocytes in HIV-infected individuals" Williams et al.J Leukocyte Biology, 2015.
- "Soluble VCAM-1 impairs human brain endothelial barrier integrity via integrin α-4-transduced outside-in signaling" Haarmann et al. Acta Neuropathology, 2015.
"Regulation of T-cell activation and migration by the kinase TBK1 during neuroinflammation" Yu et al. Nature Communications, 2015.
- "Angiotensin-(1-7) Counteracts Angiotensin II-induced Dysfunction in Cerebral Endothelial Cells via Modulating Nox2/ROS and PI3K/NO Pathways" Xiao et al. Science Direct, 2015
- "Combination treatment with AcSDKP and tissue plasminogen activator provides potent neuroprotection in rats after stroke" Zhang et al. Stroke, 2014.
- "Diabetes Mellitus Aggravates Hemorrhagic Transformation after Ischemic Stroke via Mitochondrial Defects Leading to Endothelial Apoptosis" Mishiro et al. PloS One, 2014.
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- "A systems biology analysis of brain microvascular endothelial cell lipotoxicity" Aung et al. BNC Systems Biology, 2014.
- "Downregulation of Blood-Brain Barrier Phenotype by Proinflammatory Cytokines Involves NADPH Oxidase-Dependent ROS Generation: Consequences for Interendothelial Adherens and Tight Junctions" Rochfort et al. PLoS One, 2014.
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- "Alzheimer Disease–Associated Peptide, Amyloid β40, Inhibits Vascular Regeneration With Induction of Endothelial Autophagy" Hayashi et al. Arteriosclerosis, Thrombosis, and Vascular Biology, 2009.
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- "miR-296 Regulates Growth Factor Receptor" Wurdinger et al. Cancer Cell, 2008
- "Cannabinoids inhibit HIV-1 Gp120-mediated insults in brain microvascular endothelial cells" Lu et al. J Immunology, 2008
- "Alterations in the Notch4 pathway in cerebral endothelial cells by the HIV aspartyl protease inhibitor, nelfinavir" Grigorian et al. BMC Neuroscience 2008
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- "Hypoxia-inducible factor 1 and VEGF upregulate CXCR4 in glioblastoma: implications for angiogenesis and glioma cell invasion" Zagzag et al. Laboratory Investigation, 2006
- "Integrin Regulation by Vascular Endothelial Growth Factor in Human Brain Microvascular Endothelial Cells" Lee et al. Journal of Biological Chemistry, 2006
- "CCL2/monocyte chemoattractant protein-1 mediates enhanced transmigration of human immunodeficiency virus (HIV)-infected leukocytes across the blood-brain barrier: a potential mechanism of HIV-CNS invasion and NeuroAIDS" Eugenin et al. Journal of Neuroscience, 2006
- "IL-1beta regulates blood-brain barrier permeability via reactivation of the hypoxia-angiogenesis program" Argaw et al. Journal of Immunology, 2006
- "Brain endothelial homeostasis regulation by pericytes" Kim et al. J Cerebral Blood Flow & Metabolism, 2006
- "Glioma cells under hypoxic conditions block the brain microvascular endothelial cell death induced by serum starvation" Ueda et al. J Neurochemistry, 2005
- "Human Immunodeficiency virus type 1 Nef potently induces apoptosis in primary human brain microvascular endothelial cells via the activation of caspases" Acheampong et al. Journal of Virology, 2005
- "Contribution of proteoglycans to human immunodeficiency virus type 1 brain invasion" Bobardt et al. J Virology 2004
- "Insulin degrading enzyme is expressed in the human cerebrovascular endothelium and in cultured human cerebrovascular endothelial cells" Gao et al. Neuroscience Letters, 2004
- "HIV-1 Tat-Mediated Apoptosis in Human Brain Microvascular Endothelial Cells" Avraham et al. J Immunology, 2003
- "Interactions Between p38 Mitogen-Activated Protein Kinase and Caspase-3 in Cerebral Endothelial Cell Death After Hypoxia-Reoxygenation" Lee and Lo, Stroke, 2003
- "Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis" Jiminez et al. Thrombosis Research, 2003.
- "Vascular Endothelial Growth Factor Modulates the Transendothelial Migration of MDA-MB-231 Breast Cancer Cells through Regulation of Brain Microvascular Endothelial Cell Permeability" Lee et al. J Biological Chemistry, 2003
- "mRNA expression and amino acid transport characteristics of cultured human brain microvascular endothelial cells (hBME)" Umeki et al. Drug Metabolism Pharmacokinetics, 2002
- "Vascular Endothelial Growth Factor Modulates Neutrophil Transendothelial Migration via Up-regulation of Interleukin-8 in Human Brain Microvascular Endothelial Cells" Lee et al. J Biological Chemistry, 2002
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