Mixed cells (FaDu: HF: THP-1 = 2 104:1 104:1 104) were cultured in the conventional 6-well cell culture plate (2D-4-culture) and 6-well SPL3D cell floater plate (3D-4-culture) respectively, with high-glucose DMEM medium supplemented with 10% (v/v) FBS (all from KeyGEN BioTECH). Single HUVECs were seeded in Transwell inserts at a density of 1 × 104 cells, with high-glucose DMEM medium supplemented with 10% (v/v) FBS (all from KeyGEN BioTECH). To quantify the number of migrated HUVECs, images from three different fields of view were captured and analyzed using Image J software (version 1.51j8).
Dmem high glucose medium
Manufactured by Keygen Biotech
Sourced in China, United States
DMEM high glucose medium is a commonly used cell culture media formulation that provides high levels of glucose to support the growth and proliferation of a variety of cell types. This media is optimized to maintain the viability and function of cells in in vitro culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Hyclone dmem high glucose medium, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Keygen Biotech (1 mentions)
Benzyl ether, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Beyotime (1 mentions)
Ferric acetylacetonate, by Sinopharm (1 mentions)
Methanol, by Merck Group (1 mentions)
Annexin 5 fitc pi kit, by Keygen Biotech (1 mentions)
Lysotracker, by Keygen Biotech (1 mentions)
Rhodamine phalloidin, by Keygen Biotech (1 mentions)
Hoechst 33342, by Keygen Biotech (1 mentions)
Cck 8 kit, by Keygen Biotech (1 mentions)
Fetal bovine serum (fbs), by Wisent (1 mentions)
Infinite m200, by Tecan (1 mentions)
96 well plate, by Corning (1 mentions)
8 protocols using dmem high glucose medium
1
Evaluating Migration of HUVECs in 2D and 3D
2
SH-SY5Y cell culture protocol
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SH-SY5Y cells were obtained from Jiangsu Kaiji Biotechnology Co. The cells were cultured with high glucose DMEM medium (KeyGEN, China) containing 10% fetal bovine serum (Hangzhou Sijiqing, China), 1% penicillin and streptomycin, in an incubator at 37 and with 5% CO 2 . All the cells were sub-cultured for 3 to 5 generations for experiments.
3
Synthesis and Characterization of Multifunctional Nanoparticles
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Ferric acetylacetonate (AR, Sinopharm Chemical Reagent Co. Ltd., SCRC, Shanghai, China), oleic acid (85%, Aladdin, Shanghai, China), benzyl ether (98%, Sigma-Aldrich, St. Louis, MS, USA), ethanol (AR, SCRC), n-hexane (97%, SCRC), dimercaptosuccinic acid (98%, TCI, Shanghai, China), acetone (AR, SCRC), triethylamine (AR, Aladdin, Shanghai, China), methanol (AR, SCRC), polyethyleneimine (Mw 25000, Sigma-Aldrich, St. Louis, MS, USA), heptafluorobutyric anhydride (97%, Macklin, Shanghai, China), amino-polyethylene glycol-carboxyl (Mw 2000, Xi’an Ruixi, Xi’an, China), methoxy-polyethylene glycol-polyethyleneimine (mPEG-PEI, Mw 2000-25000, Xi’an Ruixi Co. Ltd., Xi’an, China) FITC (Beyotime, Shanghai, China), CCK8 kit (KeyGen, Nanjing, China), Hoechst33342 (KeyGen, Nanjing, China), rhodamine-phalloidin (KeyGen, Nanjing, China), LysoTracker (KeyGen, Nanjing, China), Annexin V-FITC/PI kit (KeyGen, Nanjing, China), Anti-CXCR4 antibody (Abcam of Thermo Fisher Scientific Inc., Waltham, MA, USA), DMEM high glucose medium (KeyGen, Nanjing, China), fetal bovine serum (Gibco of Thermo Fisher Scientific Inc., Waltham, MA, USA), trypsin (Gibco of Thermo Fisher Scientific Inc., Waltham, MA, USA), siRNA(GenePharma Co. Ltd., Shanghai, China), 4 T1, A549, HeLa cell line was provided by Southeast University School of Medicine (Nanjing, China).
4
Transfection of Recombinant Vectors in Cell Lines
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Transfection of recombinant vectors was conducted with four cell lines: HEK-293, U87, SK-N-SH and SH-SY5Y. HEK-293 and U87 cells were cultured in HyClone® DMEM high glucose medium containing 10% fetal bovine serum (Thermo Fisher Scientific, Chelmsford, MA, USA), and the SK-N-SH cell line was treated with KeyGEN BioTECH ® DMEM high glucose medium with 0.110 g/L sodium pyruvate containing 15% fetal bovine serum. SH-SY5Y cells were cultured in Corning® DMEM/F12 1:1 mix medium +15% fetal bovine serum. All cells were cultured to a density of more than 90% at 37°C in 5% CO2 + 95% mixed air.
5
Culturing Human Cell Lines
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Human embryonic kidney HEK-293 cells were cultured in HyClone® DMEM high glucose medium containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham MA, USA), 5% CO2 + 95% mixed air at 37 °C. Neuroblastoma SK-N-SH cells were treated with KeyGEN BioTECH® DMEM high glucose medium with 0.110 g/L sodium pyruvate containing 15% fetal bovine serum under the same conditions.
6
Overexpression and Knockdown of DDX21 in LUAD Cells
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The A549 and H1299 LUAD cell line was obtained from the cell bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Cells were cultured in a DMEM high-glucose medium (KeyGEN, China) with 10% fetal bovine serum (FBS; WISENT, Canada) at 37°C with 5% CO2 (Thermo Scientific, USA). The DDX21 gene was amplified by PCR and subcloned into the pCDH513B lentiviral vector to construct a DDX21 overexpression plasmid. The virus was packaged in 293T cells, and after 48 h, a virus solution was used to infect A549 and H1299 cells. To suppress the expression of DDX21 using small interfering RNA (siRNA), cells were grown to 30%–50% confluency and transfected with siRNA targeting DDX21 or a non-specific control (NC) for 48 h. All transfections were performed using siLentFect according to the manufacturer’s instructions. siRNAs were synthesized in Genepharm, the sequences of which are listed in Supplementary Table S2A
7
Transfection of Recombinant Vectors in HEK-293 and SK-N-SH Cells
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A total of 14 recombinant vectors were transfected into human embryonic kidney cell HEK-293 and human neuroblastoma cell SK-N-SH lines (Human embryonic kidney cell HEK-293 and human neuroblastoma cell SK-N-SH were purchased from the Chinese Academy of Sciences cell bank and catalog numbers were GNHu43 and TCHu51, respectively.). HEK-293 cells were incubated in HyClone® DMEM high-glucose medium containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, Massachusetts, USA); they were maintained in 5% CO2 + 95% mixed air at 37 °C. SK-N-SH cells were treated with KeyGEN BioTECH® DMEM high-glucose medium with 0.110 g/L sodium pyruvate containing 15% fetal bovine serum under the same conditions. Transfection experiments were performed when the cell density reached 90% or more [17 (link)].
8
Evaluating Rap/Fe3O4@VHP-Lipo Cytotoxicity on Mouse Aortic Endothelial Cells
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Mouse aortic endothelial cells (MAECs) were purchased from CHI Scientific; the cells were cultured in DMEM (high glucose) medium (Jiangsu KeyGEN BioTECH Corp., Ltd., Nanjing, China) with 10% fetal bovine serum (FBS) (Gibco, South America) solution in 5% CO2 atmosphere at 37 ℃. Cell Counting Kit-8 (CCK-8, Dojindo Laboratories, Kumamoto, Japan) colorimetric assays were performed to detect cell viability of MAECs. MAECs were cultured in 96-well plates (Corning Incorporated, Corning, NY, USA) at 2 × 104 cells per well under various concentrations (0, 10, 20, 30, 40, 60 μg/mL) of Rap/Fe3O4@ VHP-Lipo for 24 h. After that, the viability of cells was estimated by measurement of absorbance at 450 nm (A450), which was read with a microplate reader (INFINITE M200, Tecan, Männedorf, Switzerland).
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