Nr8383

The alveolar macrophage cell line, NR8383, was purchased from the ATCC (USA) and maintained in Ham's F12 medium containing 10% fetal calf serum at 37°C in a humidified atmosphere containing 5% CO₂ and 95% air.
In order to evaluate the protecting effects of Rsv on LPS stimulated NR8383 cells, cells were divided into four groups: control; LPS (1 μg/mL) only; LPS (1 μg/mL) + resveratrol (40 μg/mL); resveratrol (40 μg/mL). Cells from different groups were collected for further investigation after being treated for 12 h. The concentration of LPS in this experiment was selected according to previous researches [22 (link)].
In the research of exploring the relationship between SIRT1 expression and the protecting effects of Rsv on LPS stimulated NR8383 cells, cells were divided into four groups and treated with normal Ham's F12 medium (control), LPS (1 μg/mL), LPS (1 μg/mL) + EX527 (1 μM) + resveratrol (40 μg/mL), and LPS (1 μg/mL) + resveratrol (40 μg/mL) for 12 h. Cells were then collected for further examination. By the way, EX527 was first dissolved in dimethyl sulfoxide (DMSO) to 1 mM and then diluted to the working concentration (1 μM) with Ham's F12 medium.

C. neoformans H99 was kindly provided by Assoc. Prof. Pojana Sriburee (Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand). Yeast cells were maintained on SDA and incubated at 37 °C for 72 h. A few isolated colonies were selected, cultured in SDB, incubated at 37 °C for 16–18 h, and then shaken before experimentation.
The human lung epithelial cancer cell line (A549) was kindly obtained from Asst. Prof. Dr. Khanittha Punturee (Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand) and was cultured in DMEM supplemented with 10% (v/v) fetal bovine serum (FBS), 100 units/mL of penicillin, and 100 µg/mL of streptomycin. Alveolar macrophage cell line (NR8383) (ATCC, Manassas, VA, USA) was cultured in F-12K supplemented with 15% (v/v) FBS, 100 units/mL of penicillin, and 100 µg/mL of streptomycin. The cells were maintained in a humidified atmosphere of 5% CO₂ at 37 °C.

Human lung cancer cell lines with wild-type EGFR (H358, H23, A549, H441, H69, Calu-6, and H460) (ATCC) and with EGFR mutations (H1650, H1975, HCC827, PC9, PC9-ER, and H3255) (provided by Dr. Susumu Kobayashi) were cultured in DMEM (high glucose) (Gibco) with 10% fetal bovine serum (FBS), 2 mM L-glutamine and 1% penicillin–streptomycin. Primary lung fibroblast CCD-13Lu (ATCC) and rat alveolar macrophage NR8383 (ATCC) were cultured in DMEM (high glucose) (Gibco) with 10% FBS, 2 mM l-glutamine and 1% penicillin–streptomycin. Human lung microvascular endothelial cell HULEC-5A (ATCC) was cultured in MCDB131 (Gibco) supplemented with 10 ng/ml epidermal growth factor (EGF)(Gibco), 1 μg/ml hydrocortisone (Stemcell), 10 mM l-glutamine and 10% FBS. Immortalised tracheobronchial epithelial (AALE) cells were derived as previously described and maintained in SAGM media (Lonza) [60 (link)]. Cell line identities were confirmed by STR fingerprinting and all were found negative for mycoplasma using the MycoAler Kit (Lonza).

A rat alveolar macrophage cell line (NR8383; ATCC, Manassas, VA, USA) was cultured in F12K (ATCC) with 10% FBS (Atlanta Biologicals, Lawrenceville, GA, USA) with penicillin, streptomycin, and amphotericin B (Sigma-Aldrich, St. Louis, MO, USA) at 37°C in 5% CO₂. Plating of cells with cytokine treatments was done with F12K +5% FBS. Primary alveolar macrophages were obtained by lavage from 6 week-old Fischer rats and cultured in F12K (Mediatech, Manassas, VA)+10% FBS with treatments as described below. All procedures were approved by the Institutional Animal Care and Use Committee at the Atlanta Veterans Affairs and Emory University.