human renal derived lines  (ATCC)

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: IC50s and EC50s summary table of all experiments

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Staining

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: Effect of compound exposure on cellular viability as measured by resazurin reduction. a Schematic representation of the experimental setup in RPTEC/TERT1 and HepG2 cells, the red line represents the exposure time. b Concentration response curves of resazurin reduction in RPTEC/TERT1 and HepG2 cells exposed for 24 h to a range of concentrations (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) of complex I, complex II and complex III inhibitors of the ETC. RPTEC/TERT1 (red) and HepG2 (blue). Values are represented as percentage of vehicle controls (0.1% DMSO) and further normalized to the average of at least two non-effective concentrations (if applicable) set as 100%. Measurements are average of at least three independent experiments ± SD. Connecting lines are non-linear fits ( Y = bottom + (top − bottom)/(1 + 10^((LogIC50-X) × HillSlope))) (color figure online)

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Concentration Assay

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: Oxygen consumption rates in untreated and treated RPTEC/TERT1 and HepG2 cells. Effect on key parameters of mitochondrial function measured as changes in OCR with the Seahorse analyser upon 30 min exposure to range of concentrations (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) of complex I, complex II and complex III inhibitors of the ETC in RPTEC/TERT1 and HepG2 cells. a Overview of measurable parameters after subsequent injections of test compound and modulators of the oxidative phosphorylation of the mitostress assay in HepG2 cells. Respiration is first measured at the basal level of test system (I). Decrease in OCR upon test compound injection, indicate inhibition of the mitochondrial respiration (II). Changes in OCR upon oligomycin addition, indicate the portion of oxygen employed in ATP production (III). OCR increases after the protonophore addition indicates the maximal ability of the cell to increase mitochondrial respiration (IV). Addition of antimycin A and rotenone allows for identification of non-mitochondrial respiration (V). The difference between oligomycin and rotenone/antimycin response indicates the remaining basal respiration not coupled with ATP production to be attributed to proton leakage (VI). Arrows indicate time of injections. b OCR changes after mitostress test conducted in 0.1% DMSO control samples in RPTEC/TERT1 and HepG2. Data are represented as mean of at least seven independent experiments, expressed as percentage of basal respiration ± SD. c Representative response upon exposure to rotenone (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) in HepG2 cells showing a dose dependent effect in basal (I) and maximal (II) respiration rates. d, e Plots of concentration responses in terms of oxygen consumption rates extrapolated from the mitostress test of panel compounds. Data represents the mean of two independent experiments ± SD. All measurements were normalized for basal respiration prior to compound injection, slopes are generated by plotting dose responses of the direct oxygen consumption inhibition (OCR basal, D) and inhibition of the uncoupler stimulated respiration (OCR maximal respiration, E), the latter further represented as percentage of untreated controls samples. f Response of the mitostress assay after treatment with different concentrations of cyazofamid (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M). The two highest concentrations indicate the uncoupling effect of the compound in the two cell systems

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Injection, Inhibition, Concentration Assay, Generated

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: Effect of compound exposure on mitochondrial membrane potential. Effect on mitochondrial membrane polarization by assessment of changes in mitochondrial membrane potential upon 24 h exposure to range of concentrations (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) of complex I, complex II and complex III inhibitors of the ETC in RPTEC/TERT1 and HepG2 cells. a Schematic representation of mitochondrial membrane depolarization using JC-1 and Rho123 in RPTEC/TERT1 and HepG2 respectively and representative images of changes in mitochondrial membrane polarization in RPTEC/TERT (JC-1) and HepG2 (Rho123) upon exposure to vehicle control, rotenone and antimycin A. b Concentration response curves of panel compounds in RPTEC/TERT1 (red) and HepG2 (blue). The Rho123 intensity and the intensity ratio for JC-1 were presented as percentage of 0.1% DMSO exposure. The data was further normalized to the average of at least two non-effective concentrations (if applicable). Measurements are expressed as average of at least three independent experiments ± SD (color figure online)

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Concentration Assay

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: Effect of compound exposure on extracellular lactate and extracellular acidification rates. Glycolytic switch upon decreased mitochondrial respiration was indirectly assessed by measurements of supernatant lactate and the extracellular medium acidification after 24 h exposure to range of concentrations (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) of complex I, complex II and complex III inhibitors of the ETC in RPTEC/TERT1 and HepG2 cells. a Levels of lactate in the supernatant medium. Data are represented as percentage of 0.1% DMSO controls and re-normalized to the average of at least two non-effective concentrations (if applicable) set as 100%. b Representative response to the testing concentration range of rotenone in HepG2 cells showing a dose dependent increase of medium acidification (I) reflecting the glycolytic turnover increase. c Changes in ECAR after mitostress test conducted in 0.1% DMSO control samples in RPTEC/TERT1 and HepG2. Data are represented as mean of at least seven independent experiments, expressed as percentage of basal acidification ± SD. d Plots of concentration responses of changes in ECAR after panel compounds injection. Data is mean of two independent experiments ± SD. Measurements are expressed as percentage of basal acidification and further normalized by setting the lower asymptote of the response curve to 0%, corresponding to the 100% ECAR prior to compound injection (basal acidification), and the upper asymptote to 100%, corresponding to the maximal ECAR induction (oligomycin response)

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Concentration Assay, Injection

Journal: Archives of Toxicology

Article Title: Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

doi: 10.1007/s00204-020-02792-5

Figure Lengend Snippet: Effect of medium glucose on compound induced alterations in cell viability. The effects of medium switch (glucose to galactose) in terms of cell viability was assessed in RPTEC/TERT1 and in HepG2 after 24 h exposure to a range of concentrations (1.28E−10, 6.40E−10, 3.20E−9, 1.60E−8, 8.00E−8, 4.00E−7, 2.00E−6, 1.00E−5M) of complex I, complex II and complex III inhibitors of the ETC. a Schematic representation of the carbon source switch from glucose to galactose-containing medium in RPTEC/TERT1 (5 mM Glu/Gal) and HepG2 (25 mM Glu/Gal) respectively, the red line represents the exposure time. b Plots of concentration responses in resazurin reduction. Measurements are expressed as percentage of vehicle controls (0.1% DMSO) and further normalized to the average of at least two non-effective concentrations (if applicable) set as 100%. Values are mean of two to four independent experiments ± SD. Connecting lines are non-linear fits ( Y = bottom + (top − bottom)/(1 + 10^((LogIC50-X) × HillSlope)))

Article Snippet: The human renal proximal tubule-derived cell line RPTEC/TERT1, is a non-cancerous cell line which was immortalised by introduction of the catalytic unit of human telomerase (hTERT) (Wieser et al. ).

Techniques: Concentration Assay