Chemically induced apoptotic mouse lung epithelial (MLE) cells were used for all the efferocytosis assays. MLE cells were cultured for 24 h in 6-well plates. Uniform apoptosis was induced in MLE cells (ATCC® CRL-2110™) by incubating them with 2 μM staurosporine[17 (link)]. Apoptosis was confirmed through flow-cytometry analysis after staining with Annexin V and Propidium Iodine.
Crl 2110
Manufactured by American Type Culture Collection
Sourced in United States
The CRL-2110 is a laboratory equipment product offered by the American Type Culture Collection (ATCC). It is a cell line that serves as a core function for research purposes. The product details and specifications are not available for an unbiased and concise description.
Automatically generated - may contain errors
Lab products found in correlation
Cc 4175, by Lonza (1 mentions)
Cc 3170, by Lonza (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
Crl 9609, by American Type Culture Collection (1 mentions)
Mle 12, by American Type Culture Collection (1 mentions)
Penicillin and streptomycin, by PAN Biotech (1 mentions)
Ang 1 7, by Bachem (1 mentions)
Mle 12 cells, by American Type Culture Collection (1 mentions)
Dmem f12 10, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem ham s f12, by Thermo Fisher Scientific (1 mentions)
6 protocols using crl 2110
1
Staurosporine-Induced Apoptosis in MLE Cells
2
Murine and Human Bronchial Cell Culture
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MLE-12 cells (CRL-2110, ATCC), a murine bronchial alveolar cell line, were cultured in complete DMEM. BEAS-2B cells (purchased from Procell Life Science & Technology Co. Ltd), a kind of human bronchial epithelium, were cultured in complete bronchial epithelial cell growth medium (Lonza, CC-3170 and CC-4175). The cells were incubated in a humidified atmosphere containing 95% O2 and 5% CO2 at 37°C. The cells were seeded in 24-well plates overnight and then stimulated with PBS or HDM at doses of 100 μg/mL for 24 hours.
3
Culturing Normal Lung Epithelial Cells
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Human-derived lung normal epithelial cells (BEAS-2B, CRL-9609, ATCC) and Mouse-derived lung normal epithelial cells (MLE-12, CRL-2110, ATCC) were maintained in 10% foetal bovine serum (Gibco, Grand Island, NY, USA), supplemented with 1% penicillin and 100 mg/mL streptomycin and grown at 5% CO2 and 37 °C.
4
Investigating the Effects of SiO2 and Angiotensin-(1-7) Analogs on Lung Epithelial Cells
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MLE-12 cells (mouse lung type II epithelial cell line) were obtained from the American Type Culture Collection (CRL-2110; ATCC, Manassas, VA, USA). MLE-12 cells were grown as monolayer cultures routinely in DMEM/F-12 50/50 medium (10-092-CVR; Mediatech, VA, USA) supplemented with 10% fetal bovine serum (10099141; Gibco, Thermo Fisher Scientific, Madison, WI, USA) and 1% penicillin and streptomycin (P06-07001; Pan Biotech, Aidenbach, Germany) and maintained at 5% CO2 and 37°C.
After serum starvation for 24 h, the cells were cultured with various combinations of the following: SiO2 (50 μg/cm3), 10−7 mol/L Ang-(1–7) (4084113; Bachem),19 (link) 10−5 mol/L A779, 10−4 mol/L DIZE,20 (link) and 10−5 mol/L MLN-4760.21 (link) Cells were harvested for total RNA and protein extraction after treatment for 48 h.
After serum starvation for 24 h, the cells were cultured with various combinations of the following: SiO2 (50 μg/cm3), 10−7 mol/L Ang-(1–7) (4084113; Bachem),19 (link) 10−5 mol/L A779, 10−4 mol/L DIZE,20 (link) and 10−5 mol/L MLN-4760.21 (link) Cells were harvested for total RNA and protein extraction after treatment for 48 h.
5
Murine Lung Cell Signaling Protocol
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Murine lung epithelial 12 cells (MLE) were obtained from ATCC (CRL-2110;
American Type Culture Collection, Manassas, VA) and cultured in
accordance with the manufacturer’s instructions. MLE cells were
cultured in DMEM-F12 (Gibco) with 10% FBS (DMEM-F12-10). MLE cells
were incubated with PSS (10 μM) for 1 h prior to LPS (1 μg/ml)
stimulation in an in vitro study. One h later, cells
were harvested for testing NF-κB signalling proteins by Western blot.
The total lung cells were obtained from the lung of mice according to
a protocol described before.17 (link) Lung cells were then harvested for flow cytometry.
American Type Culture Collection, Manassas, VA) and cultured in
accordance with the manufacturer’s instructions. MLE cells were
cultured in DMEM-F12 (Gibco) with 10% FBS (DMEM-F12-10). MLE cells
were incubated with PSS (10 μM) for 1 h prior to LPS (1 μg/ml)
stimulation in an in vitro study. One h later, cells
were harvested for testing NF-κB signalling proteins by Western blot.
The total lung cells were obtained from the lung of mice according to
a protocol described before.17 (link) Lung cells were then harvested for flow cytometry.
6
Fibroblast Differentiation Dynamics
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Fetal human MRC-5 lung fibroblasts (BCRC-60023, Bioresource Collection and Research Center, Hsinchu, Taiwan) and mouse MLE12 lung epithelial cells (CRL-2110, American Type Culture Collection, Manassas, VA, USA) were maintained in MEM and DMEM/Ham’s F12 supplemented with 10% FBS, 100 units/ml of penicillin, and 100 μg/ml of streptomycin (Life Technologies). MRC-5 cells were cultured in 6-well plates at a density of 2 × 105 cells/well at 37 °C in a humidified cell culture incubator with 5% CO2. Before experiments, MRC-5 cells were washed with phosphate buffered saline (PBS) and serum-starved for 24 h, prior to stimulation with HSM extract (1% or 2%, v/v) or with 2% ethanol as a control for the time indicated. TGF-β1 (5 ng/ml) was used as positive control to induce myofibroblast differentiation. At the end of the exposure period, the cells were lysed to isolate proteins for Western blot analysis.
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