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Amj2 c11

Manufactured by American Type Culture Collection
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The AMJ2-C11 is a laboratory incubator designed for controlled temperature environments. It features a stainless-steel interior, digital temperature display, and adjustable shelves. The core function of this product is to provide a stable and consistent temperature-controlled environment for various laboratory applications.

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Lab products found in correlation

Mle 12, by American Type Culture Collection (1 mentions) Svec4 10, by American Type Culture Collection (1 mentions) Recombinant il 13, by BioLegend (1 mentions) Trypan blue, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions) Pierce lal chromogenic endotoxin quantitation kit, by Thermo Fisher Scientific (1 mentions) Hbec4, by Thermo Fisher Scientific (1 mentions) Keratinocyte serum free medium, by Thermo Fisher Scientific (1 mentions)

6 protocols using amj2 c11

1

NaOCl Modulation of Allergic Inflammation

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To evaluate in vitro immunological effect of low dose NaOCl exposure aggravating allergic inflammation, we measured thymic stromal lymphopoietin (TSLP), IL-33, and IL-1β expression after treating low dose NaOCl on the epithelial and macrophage cell lines.
AMJ2-C11 (mouse macrophage cells, ATCC; CRL-2456) was used for IL-1β, caspase-1, and IL-18 expression. Cells were treated with 0.0001∼0.005% in serum free media. Cell supernatants and lysates samples were used for the analysis of mRNA and protein expression. MLE12 (mouse lung epithelial cells, ATCC; CRL-2110) was used for IL-33 expression and treated with 0.001% NaOCl up to 96 hrs. A549 (human epithelial lung carcinoma cells, American Type Culture Collection (ATCC), MD, USA; CRL-185) was used for the evaluation of TSLP. Cells were treated with 0.001% and 0.003% NaOCl for 24 hr and 48 hr and cell lysates were used for western blot.
IL-1β, IL-33 and TSLP mRNA expressions from NaOCl-treated cells were measured using real-time PCR as previously described method. Primer sequence of each gene was as follows; IL-1β, forward 5′-GCAACTGTTCCTGAACTCAACT-3′ and reverse 5′-ATCTTTTGGGGTCCGTCAACT-3′; IL-33, forward 5′- TGAGAAACCTGAAAAATGAGACCTAGA-3′ and reverse 5′- CTGCGGTGCTGCTGAACTT-3′; TSLP, forward 5′-TATGAGTGGGACCAAAAGTACCG-3′ and reverse 5′-AGTAAGGCAATGTGGCCGATT-3′.
Kim S.H., Park D.E., Lee H.S., Kang H.R, & Cho S.H. (2014). Chronic Low Dose Chlorine Exposure Aggravates Allergic Inflammation and Airway Hyperresponsiveness and Activates Inflammasome Pathway. PLoS ONE, 9(9), e106861.
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2

Culturing Murine Cell Lines for Research

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The murine endothelial cell line SVEC4-10 (CRL-2181) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). The P815 mast cells (mouse lymphoblast-like mastocytoma cell line) [66 (link),67 (link),68 (link),69 (link)] was purchased from the Rio de Janeiro Cell Bank (Rio de Janeiro, Brazil). Both cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) plus 10% heat inactivated fetal bovine serum (FBS). The murine lung macrophage cell line AMJ2-C11 (ATCC® CRL-2456™) was cultivated in DMEM plus 5% heat inactivated fetal bovine serum. All cell lines were cultured at 37 °C in a humidified environment containing 5% CO2 in air. All reagents used for cell culture were purchased from Thermo Fisher Scientific (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA).
de Souza Junior D.A., Mazucato V.M., Santana A.C., Oliver C, & Jamur M.C. (2017). Mast Cells Interact with Endothelial Cells to Accelerate In Vitro Angiogenesis. International Journal of Molecular Sciences, 18(12), 2674.
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3

Macrophage Activation by IL-13 and MSCs

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1 × 105 cells of alveolar macrophage cell line (AMJ2-C11; ATCC, Manassas, VA, USA) were seeded in a well plate. After 6 h, the cells were treated either with PBS or 20 ng/mL of recombinant IL-13 (BioLegend, San Diego, CA, USA). Naïve or Liproxstatin-1 primed-MSCs were added 12 h later. The cells were harvested after another 24 h.
Kim R.L., Bang J.Y., Kim J., Mo Y., Kim Y., Lee C.G., Elias J.A., Kim H.Y, & Kang H.R. (2022). Mesenchymal stem cells exert their anti-asthmatic effects through macrophage modulation in a murine chronic asthma model. Scientific Reports, 12, 9811.
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4

Isolation and Culture of Murine Macrophages

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Bone marrow was collected to generate bone marrow derived macrophages (BMM) and chimeric mice. Marrow was collected from femurs and tibias by rinsing and disruption through a 26-gauge needle and filtering via a 40 μM filter. BMM were differentiated in culture at 37°C using a 1:10 dilution of L929 supernatant and adherent cells were collected after one week. Alveolar macrophages were collected from exsanguinated mice by repeated lavage of the lung alveoli through a cannula with 0.6 mM EDTA in phosphate buffered saline (PBS) at 37°C. The lavage fluid was then placed on ice. Red blood cells (RBCs) were lysed using ammonium chloride/potassium bicarbonate (ACK) buffer. The murine alveolar like macrophage-cell line AMJ2-C11 was obtained from American Type Culture Collection (ATCC). Cells were counted on a hemocytometer using Trypan Blue (Sigma) to assay viability. Cells were cultured at 37°C and 5% CO2 and maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (HyClone) with 10% heat-inactivated (56°C for 30 min) fetal bovine serum (Atlanta Biologicals), 100 units penicillin, and 100 μg streptomycin (Hyclone).
Sterkel A., Mettelman R., Wüthrich M, & Klein B.S. (2015). The unappreciated intracellular lifestyle of Blastomyces dermatitidis. Journal of immunology (Baltimore, Md. : 1950), 194(4), 1796-1805.
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5

Alveolar Macrophage Exposure to PM2.5

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Mouse alveolar macrophage cells, AMJ2-C11 (American Type Culture Collection, ATCC, Rockville, MD) were cultured in DMEM supplemented with 5% fetal bovine serum (FBS), 4 mM L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 5 mM HEPES, 50 U/mL penicillin, and 50 μg/mL streptomycin, following the ATCC protocol.
Cells were seeded at 2.5 × 105 onto six-well plates and underwent a 2-h starvation at 37 °C in a 5% CO2/95% air atmosphere. Immediately following starvation, all wells were exposed to PM2.5 suspended in serum-free DMEM or controls for 3, 24, or 48 h. Control cells received media alone, media containing lipopolysaccharide (LPS) to control for potential endotoxin contamination (Tager et al. 2010 (link)), or media containing extracts of unexposed filters to control for filter material released during the extraction procedures. LPS contamination of PM2.5 samples was quantified using a chromogenic endotoxin quantitation kit (Pierce LAL Chromogenic Endotoxin Quantitation Kit; Thermo Scientific, Pittsburgh, PA) and LPS-control cells were treated with the highest LPS concentration detected from previous samples collected in Pittsburgh, PA (0.174 EU/mL). Duplicate wells for each sample/control were exposed at each time point.
Roper C., Chubb L.G., Cambal L., Tunno B., Clougherty J.E., Fattman C, & Mischler S.E. (2016). Association of IL-6 with PM2.5 Components: Importance of Characterizing Filter-Based PM2.5 Following Extraction. Water, air, and soil pollution, 228, 43.
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6

Culturing Mouse and Human Lung Cell Lines

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The mouse AM cell line, AMJ2-C11, and the mouse alveolar epithelial cell line, LA-4, were purchased from the American Type Culture Collection (Manassas, VA). The cdk4/hTERT-immortalized normal human bronchial epithelial cell line, HBEC4 [16 (link)] was obtained from the Hamon Center Collection (University of Texas Southwestern Medical Center, Dallas, TX). AMJ2-C11 and LA-4 cells were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 100 U/ml penicillin, 0.1 U/ml streptomycin, 2.5x10−4 U/ml amphotericin B, 1 mM sodium pyruvate, Minimum Essential Medium (MEM) non-essential amino acids, and 10% Fetal bovine serum (FBS). HBEC4 cells were cultured in keratinocyte serum-free medium (Life Technologies, Gaithersburg, MD) supplemented with 50 ng/ml bovine pituitary extract and 5 ng/ml epidermal growth factor. Cells were grown under standard conditions in a humidified incubator at 37°C and 5% CO2.
Takashima K., Matsushima M., Hashimoto K., Nose H., Sato M., Hashimoto N., Hasegawa Y, & Kawabe T. (2014). Protective effects of intratracheally administered quercetin on lipopolysaccharide-induced acute lung injury. Respiratory Research, 15(1), 150.
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