Human proximal tubular epithelial cells (HK-2 cells) were purchased from the Shanghai Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM (Hyclone Laboratories, Inc., Logan, UT, USA) supplemented with 10% fetal bovine serum (Biological Industries, Beit HaEmek, Israel), 100 U/ mL penicillin, and 100 μg/ mL streptomycin at 37°C and 5% CO2 according to the standard guidelines. After synchronization by culturing in DMEM for 24 h, the cells were treated with normal glucose (5.6 mM) or high glucose (30.0 mM) for 48 h, and then treated with dapagliflozin (2.5 μM, HY-10450, MedChemExpress, MCE, USA,), metformin (5 mM, Cat. NO S1950, Selleck Chemical LLC, Huston, USA), and vildagliptin (150 μM, LAF-237, Selleck Chemical LLC, USA) for 48 h in 6-well plates (Corning Inc., Corning, NY, USA).
Hy 10450
Manufactured by MedChemExpress
Sourced in United States
HY-10450 is a laboratory equipment product offered by MedChemExpress. It is a device used for the purpose of conducting scientific experiments and analyses. The core function of HY-10450 is to facilitate the measurement and manipulation of various samples or substances in a controlled laboratory environment.
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2 protocols using hy 10450
1
In Vitro Evaluation of Glucose-Lowering Drugs
2
Hypoxic Stress Response in H9c2 Cells
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H9c2 cell lines derived from rat heart were purchased from Shanghai Institute of Biochemistry and Cell Biology (C11995500BT, Shanghai, China) and were cultured in a DMEM-low glucose medium with 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc.). Normally, the cultural mediums were maintained in the condition with 5% CO2 and 95% air at 37°C (labeled as the “Normoxia group”). To induce hypoxia, the serum-free culture mediums were placed in an anoxic box (C-31, Mitsubishi GS Chemical Engineering-Plastics Co., Ltd., Shanghai, China) containing the anaerobic bag with a hypoxia indicator (C-04, Mitsubishi GS Chemical Engineering-Plastics Co., Ltd., Shanghai, China). The anaerobic system could reduce the 50% of O2 concentration within 30 min and 3 hours later, the O2 concentration would be reduced to <0.1%. To determine the optimal tolerance duration, the cell lines were incubated under hypoxic conditions for five time points along a time gradient, spanning 0 h to 48 h. The cells in either normoxia or hypoxia group needing DAPA (20μM, HY-10450, Med Chem Express) treatment should be pretreated for half an hour and then maintained in each corresponding condition, recorded as Normoxia+DAPA and Hypoxia+DAPA, respectively.
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