Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG...Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG2 cell was 3D cultured on the VitroGel system.Cell viability was detected using Cell Counting Kit-8(CCK-8) assay of HepG2 lived cell numbers.The proliferation of HepG2 cell and clustering performance was measured via fluorescence staining test.Albumin concentration in the culture medium supernatant as an index of HepG2 cell biological function was measured with ELISA kit.Independent factor tests were conducted with three key factors:inoculated cell concentration,cultured time,and dilution degree of the hydrogel.The preliminary results of independent factor tests were used to determine the levels of factors for RSM.Result The selected optimal culture conditions are as follows:concentration of inoculated cells was4.44 × 10^(5)/mL,culture time was 4.86 days,and hydrogel dilution degree was 1:2.23.The result shows that under optimal conditions,the predicted optical density(OD) value of cell viability was 3.10 and measured 2.978 with a relative error of 3.94%.Conclusion This study serves as a reference for the 3D HepG2 culture and constructs liver tissues in vitro.Additionally,it provides the foundation for repeated dose high-throughput toxicity studies and other scientific research work.展开更多
Triclosan(TCS)is widely used in personal-care products because of its bactericidal and antibacterial properties.However,TCS and its toxic byproducts have been detected in aquatic environments,animals,and plants worldw...Triclosan(TCS)is widely used in personal-care products because of its bactericidal and antibacterial properties.However,TCS and its toxic byproducts have been detected in aquatic environments,animals,and plants worldwide.TCS is a common phenolic environmental endocrine disruptor,and TCS in the environment enters the body mainly through diet and water.Most of the absorbed TCS is metabolized by the human kidneys and is eliminated in the urine[1].Some epidemiological studies have suggested a positive correlation between the TCS exposure level in urine and albumin(a biomarker of renal function),which could cause renal dysfunction[2].However,the potential effects of TCS on the mammalian kidney and its mechanisms are currently unknown.Consequently,we initiated an in vitro trial to study the molecular mechanism of cytotoxicity of TCS to glomeruli.展开更多
To explore interleukin-6(IL-6)production and characterize lipid accumulation in L02 hepatocytes induced by sodium oleate.L02 hepatocytes were incubated with 0,37.5,75,150,300,600,or 1,200μmol/L sodium oleate for 24 h...To explore interleukin-6(IL-6)production and characterize lipid accumulation in L02 hepatocytes induced by sodium oleate.L02 hepatocytes were incubated with 0,37.5,75,150,300,600,or 1,200μmol/L sodium oleate for 24 h,and the supernatant was collected to detect the concentration of IL-6.L02 hepatocytes were incubated with 300,150,75,or 0μmol/L sodium oleate for 0–24 h.The supernatant was collected for detection of IL-6 and free fatty acids.L02 hepatocytes treated with 300μmol/L sodium oleate for 0–24 h were stained with Oil Red O.With extended sodium oleate incubation time,IL-6 levels increased,and free fatty acids decreased.After 24 h incubation,IL-6 levels increased as sodium oleate increased from 37.5 to 300μmol/L(P<0.05 for 37.5μmol/L,P<0.01 for 75μmol/L and P<0.001 for concentrations 150μmol/L or higher).Lipid accumulation increased as the sodium oleate concentration and incubation time increased.Oil Red O staining intensified with incubation time extending beyond 2 h.IL-6 production and lipid accumulation in L02 hepatocytes are influenced by sodium oleate in a dose-and time-dependent manner.展开更多
基金funded by Toxicity Evaluation of Key Contaminants in Health Food by Cell-based Test Models and the Mechanism Analysis [2018YFC1602104]
文摘Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG2 cell was 3D cultured on the VitroGel system.Cell viability was detected using Cell Counting Kit-8(CCK-8) assay of HepG2 lived cell numbers.The proliferation of HepG2 cell and clustering performance was measured via fluorescence staining test.Albumin concentration in the culture medium supernatant as an index of HepG2 cell biological function was measured with ELISA kit.Independent factor tests were conducted with three key factors:inoculated cell concentration,cultured time,and dilution degree of the hydrogel.The preliminary results of independent factor tests were used to determine the levels of factors for RSM.Result The selected optimal culture conditions are as follows:concentration of inoculated cells was4.44 × 10^(5)/mL,culture time was 4.86 days,and hydrogel dilution degree was 1:2.23.The result shows that under optimal conditions,the predicted optical density(OD) value of cell viability was 3.10 and measured 2.978 with a relative error of 3.94%.Conclusion This study serves as a reference for the 3D HepG2 culture and constructs liver tissues in vitro.Additionally,it provides the foundation for repeated dose high-throughput toxicity studies and other scientific research work.
基金supported by the National 13th 5-Year Key Research grant from the Ministry of Science and Technology of the People’s Republic of China[2018YFC1602104]performed under the project"Toxicity Evaluation of Key Contaminants in Health Food by Cell-based Test Models and the Mechanism Analysis"
文摘Triclosan(TCS)is widely used in personal-care products because of its bactericidal and antibacterial properties.However,TCS and its toxic byproducts have been detected in aquatic environments,animals,and plants worldwide.TCS is a common phenolic environmental endocrine disruptor,and TCS in the environment enters the body mainly through diet and water.Most of the absorbed TCS is metabolized by the human kidneys and is eliminated in the urine[1].Some epidemiological studies have suggested a positive correlation between the TCS exposure level in urine and albumin(a biomarker of renal function),which could cause renal dysfunction[2].However,the potential effects of TCS on the mammalian kidney and its mechanisms are currently unknown.Consequently,we initiated an in vitro trial to study the molecular mechanism of cytotoxicity of TCS to glomeruli.
基金Toxicity Evaluation of Key Contaminants in Health Food by Cell-based Test Models and the Mechanism Analysis[2018YFC1602104]。
文摘To explore interleukin-6(IL-6)production and characterize lipid accumulation in L02 hepatocytes induced by sodium oleate.L02 hepatocytes were incubated with 0,37.5,75,150,300,600,or 1,200μmol/L sodium oleate for 24 h,and the supernatant was collected to detect the concentration of IL-6.L02 hepatocytes were incubated with 300,150,75,or 0μmol/L sodium oleate for 0–24 h.The supernatant was collected for detection of IL-6 and free fatty acids.L02 hepatocytes treated with 300μmol/L sodium oleate for 0–24 h were stained with Oil Red O.With extended sodium oleate incubation time,IL-6 levels increased,and free fatty acids decreased.After 24 h incubation,IL-6 levels increased as sodium oleate increased from 37.5 to 300μmol/L(P<0.05 for 37.5μmol/L,P<0.01 for 75μmol/L and P<0.001 for concentrations 150μmol/L or higher).Lipid accumulation increased as the sodium oleate concentration and incubation time increased.Oil Red O staining intensified with incubation time extending beyond 2 h.IL-6 production and lipid accumulation in L02 hepatocytes are influenced by sodium oleate in a dose-and time-dependent manner.
