Hbepc

Manufactured by Cell Applications

The HBEpC is a primary human bronchial epithelial cell line. It is derived from normal human bronchial epithelium and exhibits the typical morphological and functional characteristics of bronchial epithelial cells.

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6 protocols using hbepc

Primary Human Airway Epithelial Cell Culture

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Primary NHBE cells were obtained from Lonza and grown according to instructions. NHBE cells were cultured in T25 cell culture tissue flasks with PneumaCult-Ex Plus media (StemCell). Cells were seeded at ~100,000 cells/T25 flask and incubated at 37°C, 5% CO₂. Once cells reached 70–80% confluency, they were dissociated using 0.25% Trypsin in dissociation media and plated in 24-well transwells (Corning). Primary human bronchial epithelial cells (HBEpC) and small airway epithelial cells (HSAEpC) were obtained from Cell Applications Inc The HBEpC and HSAEpC were cultured in human bronchial/tracheal epithelial cell media and small airway epithelial cell media, respectively, following the instructions of Cell Application.
Human iPSC-derived alveolar epithelial type 2 cells (iHAEpC2) were obtained from Cell Applications Inc and cultured in growth media (i536K-05, Cell Applications Inc) according to the manufacturer’s instructions. All the primary cells were used within early passages (5–6) to avoid any gradual disintegration of the airway epithelium with columnar epithelial structure and epithelial integrity.

Tindle C., Fuller M., Fonseca A., Taheri S., Ibeawuchi S.R., Beutler N., Katkar G.D., Claire A., Castillo V., Hernandez M., Russo H., Duran J., Crotty Alexander L.E., Tipps A., Lin G., Thistlethwaite P.A., Chattopadhyay R., Rogers T.F., Sahoo D., Ghosh P, & Das S. (2021). Adult stem cell-derived complete lung organoid models emulate lung disease in COVID-19. eLife, 10, e66417.

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Culturing Human Bronchial and Carcinoma Cells

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Human Bronchial Epithelial cells HBEpC (Catalog No. 502-05a) were purchased from Cell Applications Inc., and humanderived squamous cell carcinoma SCC-25 (ATCC® CRL-1628 ™ ) was purchased from American Type Culture Collection (ATCC). HBEpC cells were cultured in bronchial/tracheal epithelial cell growth medium (Cell Applications 511-500) supplemented with 10% fetal bovine serum (FBS; Gibco 10500064), and SCC-25 cells were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F12 (DMEM/F12; Gibco 11039-021), supplemented with 10% FBS, 4 mM L-glutamine (Gibco A2916801), 1 mM sodium pyruvate (Gibco 11360070), 1× penicillin-streptomycin (equivalent to 50 U mL -1 ; Gibco 15140-122), and 400 ng mL -1 of hydrocortisone (Sigma-Aldrich H0888). All indicated values correspond to the final concentrations of the supplements. The cells were maintained in an incubator set at 37 °C and 5% CO 2 .

Mapanao A.K., Sarogni P., Santi M., Menicagli M., Gonnelli A., Zamborlin A., Ermini M.L, & Voliani V. (2022). Pro-apoptotic and size-reducing effects of protein corona-modulating nano-architectures enclosing platinum prodrug in in vivo oral carcinoma. Biomaterials science, 10(21).

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Culturing Human Bronchial and Squamous Cells

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Human bronchial epithelial cells (HBEpc) and human squamous cell carcinoma SCC-25 (HPV-negative) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). HBEpc were cultured in ready-to-use bronchia/trachea epithelial cell growth medium (Cell Applications) supplemented with 10% fetal bovine serum (FBS), while the SCC-25 cells were grown in a complete medium composed of a 1:1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s F12 (Invitrogen) supplemented with 10% FBS, 4 mM L-glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin–100 mg/mL streptomycin, and 400 ng/mL of hydrocortisone. Cells were maintained at 37 °C in a humidified incubator with 5% CO₂ atmosphere.

Madeo L.F., Sarogni P., Cirillo G., Vittorio O., Voliani V., Curcio M., Shai-Hee T., Büchner B., Mertig M, & Hampel S. (2022). Curcumin and Graphene Oxide Incorporated into Alginate Hydrogels as Versatile Devices for the Local Treatment of Squamous Cell Carcinoma. Materials, 15(5), 1648.

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Cell Culture of Human Cancer and Normal Cells

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MB231 human breast carcinoma cells, H1299 human lung carcinoma cells and H292 human lung carcinoma cells were obtained from ATCC and maintained in RPMI 1640 (Mediatec) with 10% fetal bovine serum (FBS; HyClone). Non-immortalized primary human bronchial epithelial cells (HBEpC) were obtained and maintained as suggested in bronchial/tracheal epithelial cell growth media from Cell Applications, Inc. Non-immortalized primary human mammary epithelial cells (HMEC) were obtained and maintained in mammary epithelial growth media as suggested by the vendor (Lonza). Experiments with non-transformed cells (HBEpC and HMEC) were performed between 3-10 population doublings from receipt of the cells from the company. In our experience, the doubling time of the cells did not change until after 10 population doublings. These cells maintained a healthy replicative lifespan in culture which was verified in each experiment by measuring plating efficiency.

Sciegienka S.J., Solst S.R., Falls K.C., Schoenfeld J.D., Klinger A.R., Ross N.L., Rodman S.N., Spitz D.R, & Fath M.A. (2017). D-penicillamine combined with inhibitors of hydroperoxide metabolism enhances lung and breast cancer cell responses to radiation and carboplatin via H2O2-mediated oxidative stress. Free radical biology & medicine, 108, 354-361.

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Cell Viability Assay for Cancer Cell Lines

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The cell lines U87, and A549 were obtained from ATCC, HBEpC (Human Bronchial Epithelial Cells) was obtained from Cell Applications (San Deigo, California) and MDA-231-BR was kindly donated by Patricia Steeg, NIH/NCI. The U251MG glioblastoma cell line (formerly known as U-373 MG) was originally obtained from ATCC (Manassas, VA) and was used for a maximum of fifteen passages. U87, U251MG, A549 and MDA231-BR cells were maintained in DMEM (Gibco) supplemented with 2 mM L-glutamine (Gibco) and 10% fetal bovine serum (Gibco). HBEpC were grown in bronchial/tracheal epithelial growth medium obtained from Cell Applications and were used for a maximum of three passages. All cells were maintained at 37°C and 5% CO₂. The cells were seeded into 384 well plates and allowed to grow for 24hrs and then treated as specified. After a 72 hr exposure to the indicated compounds the cells were stained with Hoescht 33342 and ethidium homodimer I for total and dead cell counts, respectively. Cells were imaged using an In Cell Analyzer 2200 and cell viability was measured based on viable nuclei count. For these studies DDC was dissolved in sterile water, DSF and Cu(DDC)₂ were solubilised in DMSO and cells were dosed such that the final DMSO concentration was 0.05%.

Wehbe M., Anantha M., Backstrom I., Leung A., Chen K., Malhotra A., Edwards K, & Bally M.B. (2016). Nanoscale Reaction Vessels Designed for Synthesis of Copper-Drug Complexes Suitable for Preclinical Development. PLoS ONE, 11(4), e0153416.

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Culturing Diverse Respiratory Cell Lines

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The stable cell line MN8CampLuc-HT-29 was cultured as described previously^{14 (link)} except addition of antibiotics and 10% FBS instead of FCS was used. The parental HT-29 cell line (ATCC: HTB-38, batch 70019050) was cultured in McCoy’s 5A (modified) medium (Life Tech., Cat. No. 26600-023) supplemented with 10% FBS (Life Tech., Cat. No. 10270-106), otherwise indicated. Human bronchial/tracheal epithelial primary cells (HBEpC) from two different donors labeled #2644 and #1865 were provided from Cell Applications (Cat. No. 502-05a). BCi-NS1.1^{35 (link)}, VA10^{36 (link)} and HBEpC cells were cultured in Bronchial/Tracheal Epithelial cell growth medium (BEGM) supplemented with retinoic acid (Cell Applications, 511A-500 and 511-RA). HBEpC were sub-cultured using subculture reagent kit (Cell Applications, 090K) and following instructions provided by the vendor. Experiments with SBI-115 antagonist were performed by culturing BCi cells in BEGM BulletKit from Lonza (Cat. No. CC-3170), as BEGM from Cell Applications was temporarily not available from the vendor. Cells were cultured at 37 °C and 5% CO₂.

Myszor I.T., Lapka K., Hermannsson K., Rekha R.S., Bergman P, & Gudmundsson G.H. (2024). Bile acid metabolites enhance expression of cathelicidin antimicrobial peptide in airway epithelium through activation of the TGR5-ERK1/2 pathway. Scientific Reports, 14, 6750.

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