Sagm small airway epithelial cell growth medium

Manufactured by Lonza

Sourced in United States

SAGM small airway epithelial cell growth medium is a complete, serum-free culture medium designed for the growth and maintenance of human small airway epithelial cells in vitro.

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Lab products found in correlation

Tgf β, by Thermo Fisher Scientific (4 mentions) Poly 1 c, by Merck Group (3 mentions) Kira8, by MedChemExpress (2 mentions) Tgfβ stimulated, by Thermo Fisher Scientific (2 mentions) Bms 345541, by Merck Group (2 mentions) Gsk126, by Selleck Chemicals (2 mentions) Hsaecs, by American Type Culture Collection (1 mentions) Compound 3, by Cayman Chemical (1 mentions) Compound 1, by Cayman Chemical (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) A549 cell, by American Type Culture Collection (1 mentions) Saecs, by Lonza (1 mentions) Saecs, by American Type Culture Collection (1 mentions) Dharmafect 1 reagent, by Horizon Discovery (1 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Cc 2547, by Lonza (1 mentions)

18 protocols using sagm small airway epithelial cell growth medium

Immortalized Human Small Airway Epithelial Cell Line

Cited 1 time

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An immortalized primary human small airway epithelial cell line was established by infecting primary hSAECs with human telomerase and cyclin dependent kinase (CDK)-4 retrovirus constructs [28 (link)]. The immortalized hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD) in a humidified atmosphere of 5% CO₂. hSAECs were stimulated with TGFβ (10 ng/mL, PeproTech, Rocky Hill, NJ) for 15 days to transformed into mesenchymal hSAECs [29 (link)]. The proteomics study had four experimental groups including normal hSAECs, TGFβ induced mesenchymal hSAECs, normal hSAECs treated with TNFα for 1 h, and mesenchymal hSAECs treated with TNFα for 1 h. Each experimental group has two biological replicates.

Zhao Y., Tian B., Sadygov R.G., Zhang Y, & Brasier A.R. (2016). Integrative proteomic analysis reveals reprograming tumor necrosis factor signaling in epithelial mesenchymal transition. Journal of proteomics, 148, 126-138.

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Immortalized hSAECs for RSV and UPR Studies

Cited 3 times

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Immortalized primary hSAECs were obtained from American Type Culture Collection (ATCC, Gaithersburg, MD, USA) and grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD, USA) in 5% CO₂ (11 (link),16 (link)). Sucrose cushion purified RSV Long strain was prepared and titered using methylcellulose plaque assay (11 (link)). hSAECs were infected at a multiplicity of infection (MOI) of 1.0 for 24 h prior to harvest. For induction of the UPR, hSAECs were treated for indicated times with various standardly used doses of 0.5–0.5 μg/ml tunicamycin (TM) or 50 nM thapsigargin (Tg). The selective IRE1α RNAse inhibitor KIRA8 (MedChemExpress, South Brunswick Township, NJ, USA) was added directly to the culture medium at a concentration of 10 μM where indicated (17 (link)).

Qiao D., Skibba M., Xu X, & Brasier A.R. (2023). Genomic targets of the IRE1-XBP1s pathway in mediating metabolic adaptation in epithelial plasticity. Nucleic Acids Research, 51(8), 3650-3670.

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Epithelial-Mesenchymal Transition in Immortalized hSAECs

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An immortalized primary human small airway epithelial cell (hSAEC) line was previously described.^{12 (link),14 (link)–18 (link)} The immortalized hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD) in a humidified atmosphere of 5% CO₂. For induction of EMT, hSAECs were TGFβ stimulated for 14 d (10 ng/mL, PeproTech, Rocky Hill, NJ).^{14 (link)}

Zhang J., Jamaluddin M., Zhang Y., Widen S.G., Sun H., Brasier A.R, & Zhao Y. (2019). Type II Epithelial-Mesenchymal Transition Upregulates Protein N‑Glycosylation To Maintain Proteostasis and Extracellular Matrix Production. Journal of proteome research, 18(9), 3447-3460.

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Immortalization of Human Small Airway Epithelial Cells

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Human small airway epithelial cells (hSAECs) were immortalized using human Telomerase/CDK4 as previously described [14 (link),15 (link)]. hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD, USA) in a humidified atmosphere of 5% CO₂. The human RSV long strain was grown in Hep-2 cells and prepared as described [7 (link),16 (link)]. The viral titer of purified RSV pools was varied from 8 to 9 log PFU/mL, determined by a methylcellulose plaque assay [17 (link)]. Viral pools were aliquoted, quick-frozen on dry ice-ethanol, and stored at −70 °C until they were used.

Xu X., Qiao D., Mann M., Garofalo R.P, & Brasier A.R. (2020). Respiratory Syncytial Virus Infection Induces Chromatin Remodeling to Activate Growth Factor and Extracellular Matrix Secretion Pathways. Viruses, 12(8), 804.

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Immortalized hSAEC Culture Protocol

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Immortalized hSAECs were previously described.^{37 (link),38 (link)} hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD) in a humidified atmosphere of 5% CO₂. Poly(I:C) was obtained from Sigma (St. Louis, MO) and used at 10 μg/mL in the cell culture. Compound 1 was purchased from Tocris and compound 3 was either purchased from Cayman Chemical (Ann Arbor, Michigan) or resynthesized in house. Compounds were dissolved in DMSO and added at the indicated concentrations.

Liu Z., Chen H., Wang P., Li Y., Wold E.A., Leonard P.G., Joseph S., Brasier A.R., Tian B, & Zhou J. (2020). Discovery of Orally Bioavailable Chromone Derivatives as Potent and Selective BRD4 Inhibitors: Scaffold Hopping, Optimization, and Pharmacological Evaluation. Journal of medicinal chemistry, 63(10), 5242-5256.

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Culturing A549 and hSAECs for Research

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Cell culture - A549 cells (human adenocarcinomic alveolar basal epithelial cells) were grown in 10% FBS - DMEM with 1% penicillin streptomycin, 1% MEM non-essential amino acids, 1% L-glutamine and 1% sodium pyruvate.^{12 (link)} An immortalized primary human small airway epithelial cell line (hSAECs) was established by infecting primary hSAECs with human telomerase and cyclin dependent kinase (CDK)-4 retrovirus constructs.^{16 (link)} The immortalized hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD). All cells were incubated at 37°C, 5% CO₂ until confluence.

Zhang Y., Sun H., Zhang J., Brasier A.R, & Zhao Y. (2017). Quantitative assessment of the effects of trypsin digestion methods on affinity purification-mass spectrometry-based protein-protein interaction analysis. Journal of proteome research, 16(8), 3068-3082.

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Culture of Human Airway Epithelial Cells

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Two types of human airway epithelial cells (HAECs)—A549 cells and primary small airway epithelial cells (SAECs)—were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured as described previously^{23 (link)}. Briefly, A549 cells were cultured in RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100 IU/mL), and streptomycin (50 μg/mL). SAECs were cultured in SAGM™ Small Airway Epithelial Cell Growth Medium (Lonza, Walkersville, MD, USA) supplemented with bovine pituitary extract, hydrocortisone, human epidermal growth factor, epinephrine, transferrin, insulin, retinoic acid, triiodothyronine, gentamicin/amphotericin-B, and bovine serum albumin. Cells were maintained at 37 °C in an atmosphere of 95% humidified air and 5% CO₂.

Choi G.S., Trinh H.K., Yang E.M., Ye Y.M., Shin Y.S., Kim S.H, & Park H.S. (2018). Role of clusterin/progranulin in toluene diisocyanate-induced occupational asthma. Experimental & Molecular Medicine, 50(5), 58.

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Immortalization and EMT Induction of Human Small Airway Epithelial Cells

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An immortalized human small airway epithelial cell (hSAEC) line was established by infecting primary hSAECs with human telomerase (hTERT) and cyclin dependent kinase (CDK)-4 retrovirus constructs [21 (link)]. The immortalized hSAECs were grown in SAGM small airway epithelial cell growth medium (Lonza, Walkersville, MD) in a humidified atmosphere of 5 % CO₂. For induction of EMT, hSAECs were TGFβ stimulated for 15 days (10 ng/ml, PeproTech, Rocky Hill, NJ). The small molecule inhibitor of IKK, BMS345541 was purchased from Sigma Aldrich and used at 10 μM [22 (link)].

Tian B., Li X., Kalita M., Widen S.G., Yang J., Bhavnani S.K., Dang B., Kudlicki A., Sinha M., Kong F., Wood T.G., Luxon B.A, & Brasier A.R. (2015). Analysis of the TGFβ-induced program in primary airway epithelial cells shows essential role of NF-κB/RelA signaling network in type II epithelial mesenchymal transition. BMC Genomics, 16(1), 529.

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Culturing Human Small Airway Epithelial Cells

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Cryopreserved human small airway epithelial cells (SAEC) were purchased from Lonza Bioscience (Alpharetta, GA) and cultured in a 37°C humidified 5% CO₂ incubator. The cells were cultured per supplier protocol using Lonza SAGM Small Airway Epithelial Cell Growth Medium and supplements. All experiments were performed between SAEC passages 1–4. For electrophysiological studies, cells were cultured on Costar Snapwell (Kennebtmk, ME) clear permeable supports with 0.4 μM polyester membrane to confluency.

Grant G.J., Coca C., Zhao X.M, & Helms M.N. (2020). Oxidized Glutathione Increases Delta-Subunit Expressing Epithelial Sodium Channel Activity in Xenopus laevis Oocytes. eMedical research, 2, 100008.

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Immortalized Human Airway Epithelial Cell Infection

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Human small airway epithelial cells (HSAECs) immortalized with human telomerase/CDK4 were obtained commercially (ATCC). These are non-oncogenic, telomerase-immortalized cells grown in the SAGM small airway epithelial cell growth medium (Lonza, cc-3118) in a humidified atmosphere of 5% CO₂. Cells were infected at confluence >90% for profiling.
The human RSV long strain was obtained from ATCC, grown in Hep-2 cells, and prepared as described (Ueba, 1978 (link)). The viral titer of purified RSV pools varied from 8 to 9 log PFU/ml, determined by a methylcellulose plaque assay (Xu et al., 2020 ). Viral pools were aliquoted and quick-frozen on dry ice–ethanol and stored at −70°C until they were used.

Xu X., Mann M., Qiao D., Li Y., Zhou J, & Brasier A.R. (2021). Bromodomain Containing Protein 4 (BRD4) Regulates Expression of its Interacting Coactivators in the Innate Response to Respiratory Syncytial Virus. Frontiers in Molecular Biosciences, 8, 728661.

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