The primary human nasal epithelial cells, HNEpCs, were purchased from PromoCell, Germany (C-12620). HNEpCs were cultured using commercially available airway epithelial cell growth medium with supplements (C-21060, PromoCell) at 37 °C, 5 % CO2. Cells were grown in tissue culture flasks coated with purified collagen (50 μg/ml) (Advanced BioMatrix). The culture medium was refreshed on every other day. The confluent monolayers of HNEpCs were washed twice with PBS before being treated with various concentrations of TMC NPs or EDIII-D3 TMC NPs (25 to 150 μg). HNEpCs cell viability was quantitated using trypan blue exclusion.
Hnepc
Manufactured by PromoCell
Sourced in Germany, United States
The HNEpC is a primary human neonatal epidermal cell line. It is a laboratory cell culture product intended for research use.
Automatically generated - may contain errors
Lab products found in correlation
Airway epithelial cell growth medium, by PromoCell (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Aecg medium, by PromoCell (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Hnepcs, by Merck Group (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Nessy 1, by Enzo Life Sciences (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Axio observer, by Zeiss (1 mentions)
Triton x 100 pbs, by Merck Group (1 mentions)
Poly l lysine coated cover glasses, by Matsunami (1 mentions)
Recombinant human leptin, by R&D Systems (1 mentions)
D lys3 ghrp 6, by Phoenix Pharmaceuticals (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Rpmi 1640 medium, by PromoCell (1 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Flagellin, by InvivoGen (1 mentions)
Il 1α, by Thermo Fisher Scientific (1 mentions)
27 protocols using hnepc
1
Evaluating Nanoparticle Impacts on Nasal Epithelial Cells
2
Quercetin's Impact on CC10 Production
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Nasal epithelial cells obtained from a healthy human’s nasal mucosa (HNEpC) (PromoCell GmbH) were suspended in AECG medium (PromoCell GmbH) at a concentration of 1 × 105 cells/mL. To examine the influence of quercetin on CC10 production from HNEpCs, cell suspensions (1.0 mL) were introduced into 24-well culture plates in triplicate containing various concentrations (1.0, 2.5, 5.0 or 7.5 μM) of quercetin and stimulated with 20.0 ng/mL of TNF [22 (link)] in a final volume of 2.0 mL. After 24 h, culture supernatants were obtained and stored at −40 °C until use.
3
In Vitro Respiratory Cell Culture
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Experiments in this study were conducted using BEAS-2B cells (RRID: CVCL_0168), A549 cells (RRID: CVCL_0023), and HNEpC. These are respiratory epithelial cell types widely used as in vitro cell culture models. BEAS-2B and A549 cell lines were purchased from Duke Cell Culture Facility (Source: ATCC, Manassas, VA) and HNEpC from PromoCell GmbH (Heidelberg, Germany). BEAS-2B cells were cultured in BEGM Bronchial Epithelial Cell Growth Medium BulletKit (Lonza, Walkersville, IL) containing BEGM basal medium and SingleQuots supplements and growth factors. Cells were plated onto flasks and plates coated with a mixture of fibronectin (0.01 mg/mL; Sigma-Aldrich, Saint Louis, MO), bovine collagen type I (0.03 mg/mL; Sigma-Aldrich) and bovine serum albumin (0.01 mg/mL; Sigma-Aldrich) dissolved in cell culture medium and overnight incubated at 37°C in a 5% CO2 and 95% humid atmosphere. A549 cells were cultured in F-12K medium supplemented with 10% fetal bovine serum and 100 units/mL penicillin and 0.1 mg/mL streptomycin. HNEpC were cultured in Airway Epithelial Cell Growth Medium (PromoCell) that contained growth supplements, growth factors and antibiotic–antimycotic mix (10 000 units/mL of penicillin, 10 000 µg/mL of streptomycin, and 25 µg/mL of Amphotericin B; ThermoFisher Scientific, Waltham, MA).
4
Cryopreserved Primary Nasal Epithelial Cells
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Cryopreserved HNEpC were purchased from PromoCell GmbH, Heidelberg, Germany (Catalog# C-1260, Lot# 436Z028). These cells are primary nasal epithelial cells obtained from the nasal mucosa of a 50-year-old Caucasian male. HNEpC were grown to 80% confluence in PromoCell’s Airway Epithelial Cell Growth Medium (Catalog# C-21060), which was supplemented with Airway Epithelial Cell Growth Medium SupplementPack (Catalog# C-39160) and Penicillin/Streptomycin.
5
Visualizing IL-33 in Human Nasal Epithelial Cells
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Human Nasal Epithelial Cells (HNEpCs) (Promocell, Rockville, MD, USA) were cultured on poly-L-lysine coated cover glasses (Matsunami, Osaka, Japan) in Airway Epithelial Cell Growth Medium (Promocell) and maintained in 5% CO2 and 95% air at 37 °C. Twelve hours after stimulation with TDI-HSA, HNEpCs were fixed with 4% sucrose-containing 4% paraformaldehyde (Sigma Chemical Co.) for 20 min. The fixed cells were permeabilized with 0.2% Triton X-100/PBS (Sigma Chemical Co.) for 5 min and blocked with 10% goat serum/PBS for 1 h. Then, an anti-IL-33 antibody (Nessy-1, 1:250, Enzo Life Sciences, Farmingdale, NY, USA) was applied overnight at 4 °C. IL-33 was visualized by isotype-specific secondary antibody conjugated with Alexa 488 (1:200, Molecular Probes, Eugene, OR, USA). A fluorescent microscope (Axio Observer, Carl Zeiss, Oberkochen, Germany) was used to obtain fluorescent images.
6
Ghrelin, GHSR1a, and Leptin Interactions
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Recombinant human ghrelin and the GHSR1a antagonist; D-Lys-3-growth hormone-releasing peptide 6 (D-Lys-3-GHRP-6) were obtained from Phoenix Pharmaceuticals. Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich. Recombinant human leptin was purchased from R&D Systems. The HNEpCs were obtained from PromoCell (C-12620).
7
Modulating miR-155-5p and SIRT1 in HNEpCs
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Primary human nasal epithelial cells (HNEpCs) were obtained from PromoCell GmbH. Cells were grown in a RPMI-1640 medium (cat. no. 31870082, Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Invitrogen; Thermo Fisher Scientific, Inc.) in a humidified incubator at 37°C with 5% CO2.
HNEpCs were cultured in a 96-well plate (3×105 cells/ml) for 24 h. miR-155-5p inhibitor (5′-ACCCCUAUCACGAUUAGCAUUAA-3′) and inhibitor control (5′-CAGUACUUUUGUGUAGUACAA-3′) were purchased from Sigma-Aldrich, Merck KGaA. The medium in the plates was replaced with a fresh one following culturing for 24 h and the HNEpCs were transfected with 50 nM miR-155-5p inhibitor and inhibitor control using Lipofectamine® 2000 transfection reagent (Thermo Fisher Scientific, Inc.). The cells were harvested at 48 h post-transfection for subsequent studies.
To explore the effect of TGF-β1 on HNEpC morphology and EMT, HNEpCs were stimulated with 5 ng/ml transforming growth factor (TGF)-β1 (cat. no. 34-8348-82; Invitrogen; Thermo Fisher Scientific, Inc.) in the presence of miR-155-5p inhibitor or inhibitor control. HNEpC morphology was observed under a DM500 light microscope (Leica Microsystems, Inc.). Furthermore, to explore the function of SIRT1, an SIRT1 inhibitor, Splitomicin (cat. no. S4068; Sigma-Aldrich; Merck KGaA) at final concentration of 100 µM was incubated with the cells for 48 h at 37°C.
HNEpCs were cultured in a 96-well plate (3×105 cells/ml) for 24 h. miR-155-5p inhibitor (5′-ACCCCUAUCACGAUUAGCAUUAA-3′) and inhibitor control (5′-CAGUACUUUUGUGUAGUACAA-3′) were purchased from Sigma-Aldrich, Merck KGaA. The medium in the plates was replaced with a fresh one following culturing for 24 h and the HNEpCs were transfected with 50 nM miR-155-5p inhibitor and inhibitor control using Lipofectamine® 2000 transfection reagent (Thermo Fisher Scientific, Inc.). The cells were harvested at 48 h post-transfection for subsequent studies.
To explore the effect of TGF-β1 on HNEpC morphology and EMT, HNEpCs were stimulated with 5 ng/ml transforming growth factor (TGF)-β1 (cat. no. 34-8348-82; Invitrogen; Thermo Fisher Scientific, Inc.) in the presence of miR-155-5p inhibitor or inhibitor control. HNEpC morphology was observed under a DM500 light microscope (Leica Microsystems, Inc.). Furthermore, to explore the function of SIRT1, an SIRT1 inhibitor, Splitomicin (cat. no. S4068; Sigma-Aldrich; Merck KGaA) at final concentration of 100 µM was incubated with the cells for 48 h at 37°C.
8
Culturing Human Nasal Epithelial Cells
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HNEpCs purchased from PromoCell GmbH (Heidelberg, Germany) were cultured under standard conditions in airway epithelial cell growth medium (PromoCell) at 37 °C in a humidified 5% CO2 incubator. For all experiments, cells of early passage numbers (six or less) were used.
9
Stimulation of Human Nasal Epithelial Cells with IL-13
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HNEpCs were purchased from PromoCell (Atlanta, GA, USA) and cultured in 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin (Thermo Fisher Scientific) in 5% CO2 atmosphere at 37°C. The cells were stimulated with Recombinant Human IL-13 (2#00-13, Escherichia coli; Peprotech, Rocky Hill, NJ, USA) at a final concentration of 5, 10, 50, 100, and 200 ng/mL for indicated time.
10
Human Nasal Epithelial Cell Stimulation
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Human nasal epithelial cells (HNEpCs) were purchased from PromoCell and grown in minimal essential medium supplemented with 10% fetal bovine serum and Antibiotic–Antimycotic (1:100, Gibco, Grand Island, NY, USA). When a confluence of 80–90% was reached, the cells were washed with PBS, and were treated with Pan3CSK4 (5 μg/mL, Invivogen), poly (I:C) (20 μg/mL, Sigma-Aldrich, St. Louis, MO), Lipopolysaccharides (LPS, 10 μg/mL, Invivogen), Flagellin (200 ng/mL, Invivogen), R848 (5 μg/mL, MedChemExpress, Monmouth Junction, NJ, USA), IFN-γ (50 ng/mL, PeproTech, Cranbury, NJ, USA), IL-4 (50 ng/mL, PeproTech), IL-5(50 ng/mL, PeproTech), IL-6 (50 ng/mL, PeproTech), IL-8 (100 ng/mL, PeproTech), IL-13 (50 ng/mL, PeproTech), indeno(1,2,3-cd)pyrene (IP, 10 nM, Sigma-Aldrich), and multiple concentrations of IL-1α (PeproTech) as noted, respectively. In some experiments, cells were pre-incubated with DEXamethasone (DEX, Sigma-Aldrich) for 2 h.
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