Hbec 5i
Manufactured by American Type Culture Collection
Sourced in United States
The HBEC-5i is a cell line derived from human bronchial epithelial cells. It is used for in vitro research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
C57 6023, by Cell Biologics (2 mentions)
Hbec 5i, by Thermo Fisher Scientific (2 mentions)
Huv ec c, by American Type Culture Collection (2 mentions)
Endothelial cell growth supplement (ecgs), by American Type Culture Collection (2 mentions)
Endothelial cell growth supplement (ecgs), by Corning (1 mentions)
Recombinant human insulin, by Merck Group (1 mentions)
M1168 kit, by Cell Biologics (1 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
Atcc htb 26, by American Type Culture Collection (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Gelatin solution, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin antibiotic solution, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Mco 18aic, by Sanyo (1 mentions)
Dmem f12, by Merck Group (1 mentions)
Rhodamine 123, by Thermo Fisher Scientific (1 mentions)
Bcecf am, by Merck Group (1 mentions)
Occludin antibody, by Thermo Fisher Scientific (1 mentions)
15 protocols using hbec 5i
1
Culturing HBMEC-60 and HBEC-5i Endothelial Cells
2
In Vitro Experiments on Brain and Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vitro experiments were conducted on human brain endothelial cells (HBEC-5i) and peripheral blood mononuclear cells (PBMC). HBEC-5i was purchased from ATCC and cultured in 10% FBS, 1% Penicillin-Streptomycin and DMEM-12F media enriched with 40 µg/mL endothelial cell growth supplement (ECGS). Culture flasks were covered with 1% gelatin. PBMCs were isolated from the buffy coat fraction from healthy blood donors obtained from Central Blood Bank and cultured in RPMI-1640 medium with 10% FBS with the addition of 1% antibiotic. Both cell lines were incubated under standard conditions (37 °C, 5% CO2).
3
MIAT and ENC1 regulation in vascular endothelial cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vascular endothelial cells HBEC-5i (ATCC® CRL-3245TM) were cultured using Dulbecco’s modified eagle’s medium (DMEM, F12, ATCC® 30-2006TM) containing 40 μg/mL of endothelial cell growth supplement and 10% fetal bovine serum (FBS). Endothelial cells were transduced separately: overexpression (oe)-negative control (NC) + short hairpin RNA (sh)-NC group (transduced with both overexpression and shRNA empty plasmids), oe-MIAT + sh-NC group (transduced with MIAT overexpression plasmid + shRNA empty plasmid), oe-NC + sh-ENC1 group (transduced with overexpression empty plasmid + shRNA-ENC1 plasmid), oe-MIAT + sh-ENC1 group (transduced with MIAT overexpression plasmid + shRNA-ENC1 plasmid).
4
Culturing Primary Mouse and Human Brain Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary C57BL/6 mouse brain microvascular endothelial cells (MBECs; C57-6023,
Cell Biologics, Chicago, IL) were seeded in 12-well plates coated with
Gelatin-Based Coating Solution (6950, Cell Biologics) for cell assays and
allowed to grow to 80–90% confluency before treatment. Cells were grown in
Complete Mouse Endothelial Cell Medium with supplemental kit (M1168 + kit, Cell
Biologics), at 37°C in 5% CO2. All experiments were completed on cell
passage 5.
Human cerebral microvascular endothelial cells were isolated from post-mortem
brains of donors and transfected with plasmid containing SV40 large T antigen
(HBEC-5i; CRL-3245TM, ATCC®, Manassas, VA). Cells were grown in F12:
DMEM plus 10% FBS and Endothelial Cell Growth Supplement (356,006, Corning) on
0.1% Gelatin (PCS-999–027, ATCC®). To exclude the effect of growth factors,
culture medium was changed to F12:DMEM without serum 4 hours prior to treatment.
All experiments were completed on cell passage 4.
All insulin stimulation and inhibitor treatments were performed at 37°C.
Hyperinsulinemic conditioning was induced in MBECs by incubating cells for
12 hours in 20 nM human recombinant insulin (I9278, Sigma, St. Louis, MO), and
HBEC-5is in 5 nM human recombinant insulin for 12 hours. Time and concentrations
for hyperinsulinemic conditioning were determined in pilot studies, data not
presented here.
Cell Biologics, Chicago, IL) were seeded in 12-well plates coated with
Gelatin-Based Coating Solution (6950, Cell Biologics) for cell assays and
allowed to grow to 80–90% confluency before treatment. Cells were grown in
Complete Mouse Endothelial Cell Medium with supplemental kit (M1168 + kit, Cell
Biologics), at 37°C in 5% CO2. All experiments were completed on cell
passage 5.
Human cerebral microvascular endothelial cells were isolated from post-mortem
brains of donors and transfected with plasmid containing SV40 large T antigen
(HBEC-5i; CRL-3245TM, ATCC®, Manassas, VA). Cells were grown in F12:
DMEM plus 10% FBS and Endothelial Cell Growth Supplement (356,006, Corning) on
0.1% Gelatin (PCS-999–027, ATCC®). To exclude the effect of growth factors,
culture medium was changed to F12:DMEM without serum 4 hours prior to treatment.
All experiments were completed on cell passage 4.
All insulin stimulation and inhibitor treatments were performed at 37°C.
Hyperinsulinemic conditioning was induced in MBECs by incubating cells for
12 hours in 20 nM human recombinant insulin (I9278, Sigma, St. Louis, MO), and
HBEC-5is in 5 nM human recombinant insulin for 12 hours. Time and concentrations
for hyperinsulinemic conditioning were determined in pilot studies, data not
presented here.
5
Culturing Human Endothelial and Breast Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cerebral microvascular endothelial cells (HBEC-5i, ATCC® CRL-3245) and Human-breast cancer cell line MDA-MB-231 (ATCC® HTB-26) were purchased from American Type Culture Collection (ATCC, United States). HBEC-5i cells were cultured as a monolayer on 0.1% gelatin solution (Gibco/Thermo Fisher, United States) coated T-flasks in DMEM:F12 medium (Gibco/Thermo Fisher, United States) supplemented with 10% FBS (Gibco/Thermo Fisher, United States), 1% penicillin/streptomycin antibiotic solution (Gibco/Thermo Fisher, United States), and 40 μg/mL endothelial growth supplement (ECGS) (Sigma-Aldrich, Spain), according to the manufacturer’s instructions. MDA-MB-231 cells were cultured as a monolayer in DMEM medium (Gibco/Thermo Fisher, United States) supplemented with 10% FBS and 1% penicillin/streptomycin antibiotic solution, according to manufacturer’s instructions. Both cells were grown in a humidified atmosphere of 5% CO2 at 37°C (MCO-18AIC (UV), Sanyo, Japan) with the medium changed every other day.
6
In Vitro Human BBB Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Astrocyte cells (human astrocytes) were from ScienCell Research Laboratory (Carlsbad, CA, USA) and endothelial cells (HBEC-5i, human brain endothelial cells) were obtained from ATCC (Manassas, VA, USA). Specific inserts for 24-well (transparent PET membrane, 0.45-µm pore diameter size) were from D. Dutscher (Alsace, France). EVOM voltohmmeter system was purchased from World Precision Instruments (Hertfondsire, UK). Occludin antibody was from Life Technologies (Saint Aubin, France), and ZO-1 was from GeneTex (San Antonio, TX, USA). Pgp, claudin-5, BCRP and MRP-1 antibodies came from Santa Cruz Biotechnology (Dallas, TX, USA). All compounds for Ringer HEPES buffer were from Sigma-Aldrich (St Quentin Fallavier, France) as well as BCECF-AM, verapamil, KO143, probenecid, Na-Fl, DMEM-F12 (Dulbecco’s Modified Eagle’s Medium) and methyl-thiazolyl-tetrazolium (MTT). Rhodamine-123 was from Thermo Fisher Scientific (Waltham, MA, USA).
7
Brain Microvascular Endothelial Cell Models
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brain microvascular endothelial cells represent acceptable in vitro models to pursue signaling studies in BBB. So, mouse brain microvascular endothelial cells (MBMEC) (C57-6023, Cell Biologics) and human brain microvascular endothelial cells (HBMEC) (HBEC-5i, ATCC) were used in our study to evaluate the effect of LPA. Cells were propagated in a growth medium containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors, and trace minerals, and supplemented with endothelial cell growth supplement, antibiotics, and fetal bovine serum. All cells were maintained at 37 °C in a humidified atmosphere of 5% CO2.
8
Brain Endothelial Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experiments were performed using immortalized human brain microvascular endothelial cells known as HBEC-5i (ATCC-CRL3245). Cells were gown in endothelial growth media (VEC technologies, Rensselaer, NY, USA) and plated on flasks coated with 0.1 % gelatin. VEC is a complete media containing FBS, antibiotics and growth factors (concentration details are not provided by manufacturer on the data sheets). Cells were grown to 80-90 % confluency and passage 5-8 were used in current experiments. Cells were serum starved in Dulbecco's Modified Eagle's medium (DMEM) for 2 h and treated with drugs in 2 % FBS containing DMEM. Cells were treated with various concentrations of ET-1 (0, 10 nM, 100 nM or 1 µM) or lipopolysaccharide (LPS, 100 ng/ml) for 24 h and viability and cell death pathways were assessed as described below and on Figure 1a. To determine the role of ET-1 in LPS-induced cell death, cells were preincubated with bosentan (10 µM) 20 min before LPS challenge. For gene expression studies, cells were harvested 6 h after stimulation.
9
Cell Line Characterization for Cancer Research
10
Culturing Human Endothelial Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human endothelial cell lines HUV-EC-C (from umbilical vein endothelium) and HBEC-5i (from cerebral microvascular endothelium) were obtained from American Type Culture Collection (ATCC, United States) and cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco, United States), 30 μg/ml endothelial cell growth supplement (E0760, Sigma) and 1% penicillin–streptomycin solution (Gibco) under hypoxic (300 μM CoCl2 for 10 h) or normoxic conditions. The cells were maintained in a humidified atmosphere incubator with 5% CO2 at 37°C and passaged regularly.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!