Wild-type human neuroblastoma cells (SK-N-SH-WT; obtained from Chongqing Pediatric Medical Research Institute, Chongqing, China) were seeded and cultured in complete medium consisting of Dulbecco's modified Eagle's medium (Beyotime, Nanjing, Jiangsu, China) with 10% FBS (Beyotime, Nanjing, Jiangsu, China) and 0.5% penicillin/streptomycin in a humidified atmosphere containing 5% CO2 at 37°C. Upon reaching 90% confluency, the cells were treated with 0.25% trypsin- ethylenediaminetetraacetic acid (EDTA; Beyotime, Nanjing, Jiangsu, China), dissociated into single-cell suspensions, and subcultured at a split ratio of 1:3 to 1:5.
Dulbecco s modified eagle s medium
Manufactured by Beyotime
Sourced in China
Dulbecco's modified Eagle's medium (DMEM) is a widely used cell culture medium formulation that provides essential nutrients and components for the growth and maintenance of various cell types. It is designed to support the optimal growth and survival of cells in in vitro culture systems. DMEM contains a balanced salt solution, amino acids, vitamins, and other essential compounds required for cell metabolism and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Beyotime (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Sk br 3, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Inh nc, by GenePharma (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mcf 7, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Beyotime (1 mentions)
Penicillin, by Beyotime (1 mentions)
Mim nc, by GenePharma (1 mentions)
Pcdna nc, by GenePharma (1 mentions)
Scramble sirna sinc, by GenePharma (1 mentions)
Matrigel, by BD (1 mentions)
Transwell plate, by Corning (1 mentions)
Inverted microscope, by Zeiss (1 mentions)
Cell counting kit 8 cck 8 assay, by Beyotime (1 mentions)
6 protocols using dulbecco s modified eagle s medium
1
Culturing Human Neuroblastoma Cells
2
Culturing Human LSCC Cell Lines
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Hep-2 and Tu177 cells (Human LSCC cell lines) were purchased from Chinese Academy of Biology, Shanghai, China. The cells were cultured in Dulbecco’s modified Eagle’s medium (Beyotime, China), supplemented with 10% fetal bovine serum (Gibco, USA). Hep-2 and Tu177 cells were incubated in a humid atmosphere (37°C and 5% CO2). The cells were passaged when cell confluence reached 70–80%. Cells were rinsed using PBS and digested using 0.25% trypsin. Lastly, the suspension of cells were centrifuged at 1000 rpm for 5 min and seeded into new plates.
3
Circular RNA regulation of breast cancer
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Human embryonic kidney cells (HEK293T), human BC cell lines (MCF7, SKBR3, MDA-MB-231) and human mammary epithelial cell line MCF-10A were available from the American Type Culture Collection (Rockville, MD, USA). These cells were cultured in Dulbecco’s modified Eagle’s medium (Beyotime, Shanghai, China) with 10% fetal bovine serum (FBS; Beyotime, Shanghai, China), 100 U/mL penicillin and 100 μg/mL streptomycin (Beyotime, Shanghai, China) at 37°C in 5% CO2. Circ_0048764 overexpression plasmid (pcDNA-circ_0048764), empty vector cDNA (pcDNA-NC), small interfering RNAs (siRNAs) targeting circ_0048764 (si-circ_0048764-1 and si-circ_0048764-2), siRNA negative control (scramble siRNA, si-NC), miR-1296-5p mimics and its control (mim-NC), miR-1296-5p inhibitors and its control (Inh-NC) were produced by GenePharma (Shanghai, China). The above vectors were subsequently transfected into SKBR3 and MCF7 cells by Lipofectamine®3000 (Invitrogen, Carlsbad, CA, USA). 24 h later, quantitative real-time polymerase chain reaction (qRT-PCR) was executed to detect the efficacy.
4
Transwell Invasion Assay for Cell Migration
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Transwell assays were conducted to analyze the migration and invasion abilities of cells. Briefly, 4×104 cells in 100 µl RPMI-1640 culture medium supplemented with 10% FBS were placed in the upper chamber of Transwell plates (Corning Inc., Corning, NY, USA), which were pre-coated with Matrigel (growth factor reduced; BD Biosciences). Dulbecco's modified Eagle's medium (Beyotime Institute of Biotechnology) with 20% FBS was added to the lower chambers. After 24-h incubation, cells that had invaded across the membrane were fixed in 95% methanol at room temperature and stained with 0.1% crystal violet for 10 min at room temperature. Cells in nine randomly selected fields were counted under an inverted microscope (Carl Zeiss AG, Jena, Germany) at ×200 magnification, and the number of invading cells was expressed as the mean cell number in these fields.
5
Cell Proliferation Assay for A549 and H358 Lung Cancer Cells
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A549 and H538 were purchased from the Chinese Academy of Science Cell Bank and cultured in Dulbecco's Modified Eagle's Medium (DMEM Beyotime, CN) supplemented with 10% Fetal bovine serum (FBS, EveryGreen, Zhejiang, China) and 1% antibiotics at 37 °C in 5% CO2. Small interfering RNA (siRNA) and the corresponding negative control (siNC) were purchased from Ribobio (Shanghai, China). A549 and H358 cells were transfected with siDARS2, siCOX5B, and siNC using the Lipo8000 Transfection Reagent (Beyotime Biotechnology, China). A549 cells with GFP fluorescence were obtained as reported previously [29 (link)]. Cell viability assay was also performed using Cell Counting Kit -8 (CCK-8) assay (Beyotime, CN) according to the previous study. The relative cell viability was calculated as follows: Relative Cell Viability = (Test absorbance/Mean absorbance of control wells). At the logarithmic growth phase, 1500 cells were seeded into 24-well plates (Corning, NY, USA) at 100 μL of cell suspension per well. Cell proliferation was measured according to corresponding fluorescence intensity after incubation for 0, 24, 48, 72, 96, and 120 h at 37 °C. Images were acquired on the Opera Phenix High Content Screening System (PerkinElmer) and analyzed on Harmony High-Content Imaging and Analysis Software.
6
Silencing SiRT1 in HepG2 and MHCC97H Cells
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Cells and cell culture. HepG2 and MHCC97H cells were procured from the American Type Culture Collection (USA). They were cultured in Dulbecco's modified Eagle's medium (Beyotime, China) containing 10% fetal bovine serum (Gibco, USA).
The cells were then incubated at 37°C in a humid atmosphere (5% CO 2 ) and either passaged or used for further experiments when cell confluence reached 70-80%.
Cell transfection. A small interfering (si) RNA targeting SiRT1 (si-SiRT1) and a negative control siRNA (si-NC) were purchased from GenePharma (China). Lipofectamine 3000 (Invitrogen, USA) was used as the transfection reagent, and transfection was performed based on the manufacturer's protocols.
The cells were then incubated at 37°C in a humid atmosphere (5% CO 2 ) and either passaged or used for further experiments when cell confluence reached 70-80%.
Cell transfection. A small interfering (si) RNA targeting SiRT1 (si-SiRT1) and a negative control siRNA (si-NC) were purchased from GenePharma (China). Lipofectamine 3000 (Invitrogen, USA) was used as the transfection reagent, and transfection was performed based on the manufacturer's protocols.
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