Human cervical carcinoma HeLa cell line, human lung carcinoma A549 cell line and human umbilical vein endothelial HUVEC cell line were obtained from KeyGEN Biotech Co. Ltd. (Nanjing, China). All cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS, 100 μg mL-1 streptomycin and 100 U mL-1 penicillin at 37 °C in a humidified incubator containing 5% CO2 and 95% air. The medium was replenished every other day and the cells were subcultured after reaching confluence. Cell numbers were determined with a Petroff-Hausser cell counter (USA).
A549 cell line
Manufactured by Keygen Biotech
Sourced in China
The A549 cell line is a type of human alveolar basal epithelial cell line derived from a lung carcinoma. It is commonly used in research to study various aspects of lung biology and disease, including cell signaling, metabolism, and response to treatments.
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4 protocols using a549 cell line
1
Cell Culture Conditions for Cancer and HUVEC
2
Cell Culture of Human Lung Lines
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The human pulmonary epithelial A549 cell line was obtained from KeyGen Biotech (Nanjing, China). The human fetal lung fibroblast cell line HFL1 was kindly supplied by Professor Wen Ning (Nankai University). The cells were cultured in a medium supplemented with 10% heat-inactivated (56 °C, 30 min) fetal calf serum (HyClone, USA) and maintained at 37 °C with 5% CO2 in a humidified atmosphere.
3
Cytotoxicity and Uptake of CN-NSs
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The Beas-2B cell line was obtained from KeyGEN BioTECH Co., Ltd. (Nanjing, China). The cell was cultured in a BEBM medium contain 90% RPMI-1640 and 10% FBS, and incubated at 37 °C with 5% CO2. The Beas-2B cells were firstly seeded in 96-well plates (1 × 105 cells/mL) and cultured for 24 h before drug treatment. Then, the different concentrations of CN-NS were dropped into the cell wells. After treatment for 24 h, the cells were incubated with 10 μL of CCK8 (KeyGEN BioTECH, Nanjing, China) at 37 °C for 4 h. Then, the optical density was measured at 450 nm with a microplate plate reader (BioTek ELx800, Winooski, VT, USA).
The A549 cell line was obtained from KeyGEN BioTECH Co., Ltd. (Nanjing, China). The A549 cell was cultured in a medium contain 90% F-12K and 10% FBS. The cells were incubated at 37 °C with 5% CO2. The cellular uptake evaluation of CN-NSs was determined using flow cytometer. After culturing for 2 days, the A549 cells were treated with 1 mL CN-NSs (50 μg/mL) at 37 °C for 2 h. Additionally, the culture medium was removed, the cells were washed 2–3 times with PBS, trypsinized and centrifuged. The supernatant was removed, and the centrifuged cells were gently resuspended in 0.25 mL PBS. The mean fluorescence intensity of cells was determined using the flow cytometer.
The A549 cell line was obtained from KeyGEN BioTECH Co., Ltd. (Nanjing, China). The A549 cell was cultured in a medium contain 90% F-12K and 10% FBS. The cells were incubated at 37 °C with 5% CO2. The cellular uptake evaluation of CN-NSs was determined using flow cytometer. After culturing for 2 days, the A549 cells were treated with 1 mL CN-NSs (50 μg/mL) at 37 °C for 2 h. Additionally, the culture medium was removed, the cells were washed 2–3 times with PBS, trypsinized and centrifuged. The supernatant was removed, and the centrifuged cells were gently resuspended in 0.25 mL PBS. The mean fluorescence intensity of cells was determined using the flow cytometer.
4
Nude Mice for Lung Cancer Xenograft
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All animal care, use, and experimental procedures were approved by the Animal Affairs Committee of Jiangsu Academy of Traditional Chinese Medicine (Nanjing, Jiangsu, People's Republic of China). BALB/c nu/nu male nude mice (18–20 g), 4 to 6 weeks old, were provided by the Experimental Animal Center of the same agency. Human non-small cell lung cancer A549 cell line was obtained from KeyGen Biotech Co. Ltd (Nanjing, China), was cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS, GIBCO), 100 units/ml penicillin, and 100 mg/ml of streptomycin at 37°C in 5% CO2. Xenografts were initiated by inoculating 1 × 106 tumor cells subcutaneously into a unilateral flank region of mice. All animals were given Lugol's solution in drinking water to block thyroid gland from taking up free 131I.
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