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Human Bronchial Epithelial Cells: HBEpC

(Click to Enlarge) Top: HBEpC culture at low magnification (A) and immunolabeled for cytokeratin 18 (B). Bottom: Characterization of the HBEpC based 3D airway tissue model, showing Millipore Insert, day 14 (A).  High resolution confocal imaging (B–G) of the top cell layer (B,D,F) or a cross section (C,E,G).  Shown are DAPI-labeled Nuclei (B&C), rhod-phall-labeled Actin (D&E) and merged images (F&G).

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HBEpC provide an excellent model system to study all aspects of epithelial function and disease, particularly those related to airway viral infections, as well as tissue repair mechanisms, signaling changes and potential treatments relevant to lung injuries, mechanical and oxidative stress, inflammation, pulmonary diseases and smoking.  Note that Cell Applications, Inc. offers HTEpC obtained from healthy donors, as well as from asthma patients.

When grown on inserts and provided with the liquid/air interface, HBEpC can differentiate into a pseudostriated epithelium and serve as a more physiological 3D tissue model for in vitro studies (see Figure on the left).

HBEpC from Cell Applications, Inc. have been used to:

  • provide a control to evaluate cytokine secretion in a cell line A549, demonstrating increased GM-CSF and IL-8 secretion in response to TNF-alpha or IL-1beta stimulation and determine that immortalized cells are a good fit for primary assays, but the results should be confirmed using primary cells (Lee, 2005);
  • demonstrate that mechanical strain of primary bronchial airway epithelial cells induced Erk1/2 activation leading to upregulation of NO synthase and PlGF, a member of the VEGF family (Mohammed, 2007); 
  • show that lung injury alters the localization of hepatocyte growth factor activator inhibitor type 1 (Tanaka, 2009); and upregulates HGF leading to increased COX-2 expression via β-catenin, Akt, and p42/p44 MAPK dependent pathway, contributing to tissue repair through activating cell proliferation (Lee, 2008);
  • investigate the mechanism of angiotensin II (Ang-II) induced apoptosis, and demonstrate that Ang-II-activates SHP-2 that inhibits nucleolin binding to Bcl-x(L) mRNA, lowering levels of the anti-apoptotic Bcl-2 protein family members (Lee, 2010);
  • demonstrate that pulmonary fibrosis caused by bleomycin, is mediated via Fas and caspase-8, -3 and -6 dependent apoptotic pathway (Mungunsukh, 2010);
  • show that Pyocyanin, a P. aeruginosa toxin, increases oxidative stress in epithelial cells by inhibiting their catalase activity (O’Malley, 2003) and also increases production of IL-8 and ICAM-1 (Look, 2005);
  • demonstrate that airway edema caused by RSV infection results from NO-dependent HIF-1α stabilization and VEGF production leading to increased bronchial epithelial permeability (Kilani, 2004a, b);
  • study the mechanisms of viral entry (Wan, 2007; Song, 2009) and associated pro-inflammatory signaling (Othumpangat, 2012a); demonstrate that defensins prevent viral fusion and cell entry by a general mechanism that involves crosslinking membrane glycoproteins (Leikina, 2005); and evaluate the ability of DAS181 (Fludase™), a sialidase fusion protein, to protect against viral infection (Triana-Baltzer, 2009);
  • investigate the role of Myc in HPV-induced cancerogenesis (Narisawa-Saito, 2012) and identify DLG4 as a tumor suppressor targeted by E6 during HPV infection (Handa, 2007); and identify the gene for c-myc promoter-binding protein-1 (MBP-1), a transcriptional tumor suppressor (Lung, 2010);
  • demonstrate that RSV upregulates the NGF-TrKA signaling in human airways by silencing miR-221 expression, and this favors viral replication by preventing apoptosis (Othumpangat, 2012b).
  • investigate the mechanisms of chronic obstructive pulmonary disease by demonstrating that endothelin-1 downregulates ACE2 expression via the ETA receptor and p38 MAPK-dependent mechanism (Zhang, 2013); and study the anti-inflammatory action of therapeutic drugs fluticasone propionate and salmeterol (Nasreem, 2012, 2014);
  • show that Pen ch 13, major Penicillium allergen, down-regulates CD44 protein expression in airway epithelial cells, contributing to lung inflammation and prolonging the repair response (Tai, 2010);
  • find that mir-218 mediates smoking-induced gene expression changes in bronchial epithelia (Schembri, 2009).
  • provide, along with Human Epidermal Keratinocytes, also from Cell Applications, Inc., control RNA samples to assess expression of thymidylate synthase and demonstrate its 79-fold increase in cancer cells, compared to normal primary cells (Shirasaki, 2008);
  • investigate, along with Human Hair Follicle Dermal Papilla Cells and Human Epidermal Keratinocytes, all from Cell Applications, Inc., mechanisms of cellular senescence, and develop methodology to extend cellular life span and immortalize the cells by inhibiting p16INK4a and introducing hTERT (Haga, 2007);
  • develop a BioMAP assay platform and screen EPA ToxCast library compounds to characterize effects relevant to human tissue and inflammatory disease biology (Houck, 2009; Berg, 2010), as well as compounds relevant to treatment of autoimmune inflammatory diseases (Haselmayer et al, 2014);
  • develop an in vitro assay to study mucin secretion (Abdullah, 2012).

Characterization: the cells have a characteristic morphology consistent with an epithelial origin and are positive for epithelial cell marker cytokeratin 18.

Tissue:
Surface epithelium of normal bronchi. Each lot tested negative for HIV, Hepatitis B and Hepatitis C and negative for mycoplasma, bacteria, yeast, fungi.
Cryopreserved ampoule:
1st passage, >500,000 cells in Cell Basal Medium containing 10% FBS & 10% DMSO.
Kit contains:
Ampoule of cryopreserved Human Bronchial Epithelial Cells (502-05a), 500 ml of Bronchial/Tracheal Epithelial Cell Growth Medium (511-500), (511-500), and a Subculture Reagent Kit (090K).
Proliferating Cells:
 
Shipped in Growth Medium at 2nd passage in either flasks or multiwell dishes.
Population doublings:
Can be cultured at least 16 doublings. 

2015
Burns, T., A. Ali, and D. Matesic. 2015. Comparative Effects of 4-Phenyl-3-Butenoic Acid and Vorinostat on Cell Growth and Signaling. Anticancer Research, 35: 775-784.
2014
Haselmayer, P., M. Camps, M. Muzerelle, S. El_bawab, C. Waltzinger, L. Bruns, N. Abla, M. Polokoff, C. Jond-necand, and M. Gaudet. 2014. Characterization of novel PI3Kδ inhibitors as potential therapeutics for SLE and lupus nephritis in pre-clinical studies. Name: Frontiers in Immunology. 5:233.
Nasreen, N., L. Gonzalves, S. Peruvemba, and K.A. Mohammed. 2014. Fluticasone furoate is more effective than mometasone furoate in restoring tobacco smoke inhibited SOCS-3 expression in airway epithelial cells. International Immunopharmacology. 19:153-160.
2012
Abdullah, L., C. Wolber, M. Kesimer, J. Sheehan, and C.W. Davis. 2012. Studying Mucin Secretion from Human Bronchial Epithelial Cell Primary Cultures. In Mucins. Vol. 842. M.A. McGuckin and D.J. Thornton, editors. Humana Press. 259-277.
Narisawa-Saito, M., Y. Inagawa, Y. Yoshimatsu, K. Haga, K. Tanaka, N. Egawa, S.-i. Ohno, H. Ichikawa, T. Yugawa, M. Fujita, and T. Kiyono. 2012. A critical role of MYC for transformation of human cells by HPV16 E6E7 and oncogenic HRAS. Carcinogenesis. 33:910-917.
Nasreen, N., N. Khodayari, B. Sukka-Ganesh, S. Peruvemba, and K.A. Mohammed. 2012. Fluticasone propionate and Salmeterol combination induces SOCS-3 expression in airway epithelial cells. International Immunopharmacology. 12:217-225.
Othumpangat, S., M. Regier, and G. Piedimonte. 2012a. Nerve growth factor modulates human rhinovirus infection in airway epithelial cells by controlling ICAM-1 expression. American Journal of Physiology. 302:L1057-L1066.
Othumpangat, S., C. Walton, and G. Piedimonte. 2012b. MicroRNA-221 Modulates RSV Replication in Human Bronchial Epithelium by Targeting NGF Expression. PloS one. 7:e30030.
2010

Berg, E.L., J. Yang, J. Melrose, D. Nguyen, S. Privat, E. Rosler, E.J. Kunkel, and S. Ekins. 2010. Chemical target and pathway toxicity mechanisms defined in primary human cell systems. Journal of Pharmacological and Toxicological Methods. 61:3-15.

Lung, J., K.-J. Liu, J.-Y. Chang, S.-J. Leu, and N.-Y. Shih. 2010. MBP-1 is efficiently encoded by an alternative transcript of the ENO1 gene but post-translationally regulated by proteasome-dependent protein turnover. FEBS Journal. 277:4308-4321.

Maier, K.G., X. Han, B. Sadowitz, K.L. Gentile, F.A. Middleton, and V. Gahtan. 2010. Thrombospondin-1: a proatherosclerotic protein augmented by hyperglycemia. Journal of Vascular Surgery. 51:1238-1247.

Mungunsukh, O., A.J. Griffin, Y.H. Lee, and R.M. Day. 2010. Bleomycin induces the extrinsic apoptotic pathway in pulmonary endothelial cells. American Journal of Physiology - Lung Cellular and Molecular Physiology. 298:L696-L703.

Tai, H.Y., M.F. Tam, H. Chou, D.W. Perng, and H.D. Shen. 2010. Pen ch 13 Major Fungal Allergen Decreases CD44 Expression in Human Bronchial Epithelial Cells. International Archives of Allergy and Immunology. 153:367-371.

2009
Houck, K.A., D.J. Dix, R.S. Judson, R.J. Kavlock, J. Yang, and E.L. Berg. 2009. Profiling Bioactivity of the ToxCast Chemical Library Using BioMAP Primary Human Cell Systems. Journal of biomolecular screening. 14:1054-1066.
Schembri, F., S. Sridhar, C. Perdomo, A.M. Gustafson, X. Zhang, A. Ergun, J. Lu, G. Liu, X. Zhang, J. Bowers, C. Vaziri, K. Ott, K. Sensinger, J.J. Collins, J.S. Brody, R. Getts, M.E. Lenburg, and A. Spira. 2009. MicroRNAs as modulators of smoking-induced gene expression changes in human airway epithelium. PNAS. doi: 10.1073/pnas.0806383106.
Song, H., H. Wan, Y. Araya, and D. Perez. 2009. Partial direct contact transmission in ferrets of a mallard H7N3 influenza virus with typical avian-like receptor specificity. Virology Journal. 6:126.
Tanaka, H., T. Fukushima, K. Yorita, M. Kawaguchi, and H. Kataoka. 2009. Tissue injury alters the site of expression of hepatocyte growth factor activator inhibitor type 1 in bronchial epithelial cells. Human Cell. 22:11-17.
Tumpey, and F. Fang. 2009. Novel Pandemic Influenza A(H1N1) Viruses Are Potently Inhibited by DAS181, a Sialidase Fusion Protein. PloS one. 4:e7788.
2010
Berg, E.L., J. Yang, J. Melrose, D. Nguyen, S. Privat, E. Rosler, E.J. Kunkel, and S. Ekins. 2010. Chemical target and pathway toxicity mechanisms defined in primary human cell systems. Journal of Pharmacological and Toxicological Methods. 61:3-15.
Lee, Y.H., O. Mungunsukh, R.L. Tutino, A.P. Marquez, and R.M. Day. 2010. Angiotensin-II-induced apoptosis requires regulation of nucleolin and Bcl-xL by SHP-2 in primary lung endothelial cells. Journal of cell science. 123:1634-1643.
Lung, J., K.-J. Liu, J.-Y. Chang, S.-J. Leu, and N.-Y. Shih. 2010. MBP-1 is efficiently encoded by an alternative transcript of the ENO1 gene but post-translationally regulated by proteasome-dependent protein turnover. FEBS Journal. 277:4308-4321.
Mungunsukh, O., A.J. Griffin, Y.H. Lee, and R.M. Day. 2010. Bleomycin induces the extrinsic apoptotic pathway in pulmonary endothelial cells. American Journal of Physiology - Lung Cellular and Molecular Physiology. 298:L696-L703.
Tai, H.Y., M.F. Tam, H. Chou, D.W. Perng, and H.D. Shen. 2010. Pen ch 13 Major Fungal Allergen Decreases CD44 Expression in Human Bronchial Epithelial Cells. International Archives of Allergy and Immunology. 153:367-371.
2008

Lee, Y.H., Y.J. Suzuki, A.J. Griffin, and R.M. Day. 2008. Hepatocyte growth factor regulates cyclooxygenase-2 expression via β-catenin, Akt, and p42/p44 MAPK in human bronchial epithelial cells. American Journal of Physiology. 294:L778-L786.

2007

Haga, K., S.-i. Ohno, T. Yugawa, M. Narisawa-Saito, M. Fujita, M. Sakamoto, D.A. Galloway, and T. Kiyono. 2007. Efficient immortalization of primary human cells by p16INK4a-specific short hairpin RNA or Bmi-1, combined with introduction of hTERT. Cancer science. 98:147-154.
Handa, K., T. Yugawa, M. Narisawa-Saito, S.-i. Ohno, M. Fujita, and T. Kiyono. 2007. E6AP-Dependent Degradation of DLG4/PSD95 by High-Risk Human Papillomavirus Type 18 E6 Protein. Journal of Virology. 81:1379-1389.
Horvath, G., E.S. Mendes, N. Schmid, A. Schmid, G.E. Conner, M. Salathe, and A. Wanner. 2007. The effect of corticosteroids on the disposal of long-acting β2-agonists by airway smooth muscle cells. Journal of Allergy and Clinical Immunology. 120:1103-1109.
2006
Tai, H., M. Tam, H. Chou, H. Peng, S. Su, D. Perng, and H. Shen. 2006. Pen ch 13 allergen induces secretion of mediators and degradation of occludin protein of human lung epithelial cells. 61:382-388.

2005

Lee, S.C., J.Y. Hsu, L.S. Fu, J.J. Chu, S.J. Fan, and C.S. Chi. 2005. Comparison of the activities of granulocyte-macrophage colony-stimulating factor and interleukin-8 secretion between two lung epithelial cell lines. J. microbial., immunol.& infection. 38:327-331.

Look, D.C., L.L. Stoll, S.A. Romig, A. Humlicek, B.E. Britigan, and G.M. Denning. 2005. Pyocyanin and Its Precursor Phenazine-1-Carboxylic Acid Increase IL-8 and Intercellular Adhesion Molecule-1 Expression in Human Airway Epithelial Cells by Oxidant-Dependent Mechanisms. The Journal of Immunology. 175:4017-4023.

2004

Kilani, M., K. Mohammed, N. Nasreen, R. Tepper, and V. Antony. 2004a. RSV Causes HIF-1α Stabilization via NO Release in Primary Bronchial Epithelial Cells. Inflammation. 28:245-251.

Kilani, M.M., K.A. Mohammed, N. Nasreen, J.A. Hardwick, M.H. Kaplan, R.S. Tepper, and V.B. Antony. 2004b. Respiratory syncytial virus causes increased bronchial epithelial permeability*. CHEST Journal. 126:186-191.

2003
O'Malley, Y.Q., K.J. Reszka, G.T. Rasmussen, M.Y. Abdalla, G.M. Denning, and B.E. Britigan. 2003. The Pseudomonas secretory product pyocyanin inhibits catalase activity in human lung epithelial cells. American Journal of Physiology. 285:L1077-L1086.

 

Products

3-D Airway Model

Pricing
Description
Size
Cat. #
Price
3-D Airway Model 3D Human Bronchial Epithelial Cells (HBEpC), pre-grown and differentiated into pseudostriated epithelium on PCF inserts. 24 Inserts 502-3D-24 $1,250.00
3-D Airway Model 3D Human Bronchial Epithelial Cells (HBEpC), pre-grown and differentiated into pseudostriated epithelium on PCF inserts. 12 Inserts 502-3D-12 $705.00

3-D Airway Model Kit

Pricing
Description
Size
Cat. #
Price
3-D Airway Model Kit Cryopreserved HBEpC, media, twelve PCF inserts, reagents, subculture kit, and detailed instructions, to form a 3-D Airway Model. 1 Kit 502K-3D $705.00

Cryopreserved HBEpC, adult

Pricing
Description
Size
Cat. #
Price
Cryopreserved HBEpC, adult Each ampoule contains >500,000 cryopreserved cells. Each kit contains one ampoule, 500 ml Growth Medium (511-500), and a Subculture Reagent Kit (090K). 1 Kit 502K-05a $795.00
Cryopreserved HBEpC, adult 1 Ampoule 502-05a $646.00

Proliferating HBEpC, adult

Pricing
Description
Size
Cat. #
Price
Proliferating HBEpC, adult Cultured and shipped at second passage in Bronchial/Tracheal Epithelial Cell Growth Medium. T-25 Flask 503-25a $646.00
Proliferating HBEpC, adult T-75 Flask 503-75a $836.00
Proliferating HBEpC, adult 6 well 503-6Wa $836.00
Proliferating HBEpC, adult 96 well 503-96Wa $956.00

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