sodium glycocholate (gca)  (Nacalai)

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Effect of different bile acids on HuSaV replication efficiency in Caco‐2/Cas9 cells. Caco‐2/Cas9 cells were inoculated with HuSaV GI.1 (AH20)‐positive stool suspension (approximately 4 × 10 7 copies/well in 96‐well plates) and incubated at 37°C for 3 h. HuSaV RNA copy numbers in the culture supernatants were measured at 0 dpi (immediately after medium replacement) and 7 dpi with different bile acids in the culture media. Each dot represents individual data points; bars indicate the geometric mean HuSaV RNA copy numbers; error bars represent the geometric SD. This experiment was performed once with five technical replicates. GCA, Sodium glycocholate; GCDCA, sodium glycochenodeoxycholate.

Article Snippet: In accordance with a previous study, 1000 μM sodium glycocholate (GCA; Nacalai Tesque, Kyoto, Japan) was added in the infection assay (Takagi et al. ).

Techniques: Suspension, Incubation

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Effect of different bile acids on HuSaV replication efficiency in Caco‐2MC cells. Caco‐2MC cells were inoculated with HuSaV GI.1 (M13‐18) stock at an MOI of 0.01 and incubated at 37°C for 3 h. HuSaV RNA copy numbers in the cell lysates were measured at 0 dpi (after 3 h incubation) and 5 dpi with different bile acids in the culture media. Each dot represents individual data points; bars indicate the geometric mean HuSaV RNA copy numbers; error bars represent the geometric SD. This experiment was performed three times with five technical replicates. GCA, sodium glycocholate; GCDCA, sodium glycochenodeoxycholate.

Article Snippet: In accordance with a previous study, 1000 μM sodium glycocholate (GCA; Nacalai Tesque, Kyoto, Japan) was added in the infection assay (Takagi et al. ).

Techniques: Incubation

Journal: Foods

Article Title: Effect of Coffee on the Bioavailability of Sterols

doi: 10.3390/foods11192935

Figure Lengend Snippet: Amount of LY transported and corresponding permeability coefficient in permeability assays through Caco-2 monolayers with a multi-time point assay with sampling at 2, 4 and 6 h. Effect of DHE (10 µM) emulsified by GCA (10 mM), in the absence and presence of coffee extracts, n = 2.

Article Snippet: Dulbecco’s modified Eagle’s medium high glucose (DMEM, 4.5 g/L glucose with 2 mM L-glutamine), sodium bicarbonate, nonessential amino acids (NEAA), penicillin (10,000 units/mL)–streptomycin (10 mg/mL) antibiotic, 0.25% ( w / v ) trypsin, ethylenediamine tetraacetic acid (EDTA), Hank’s balanced salt solution (HBSS), 4-2-hydroxyethyl-1-piperazineethanesulfonic acid (HEPES), Lucifer yellow CH di-potassium salt (LY), Hoechst 33,342, dehydroergosterol (DHE) and the sodium salts of glycocholic acid (GCA), glycodeoxycholic acid (GDCA) and glycochenodeoxycholic acid (GCDCA) were all purchased from Merck S.A (Algés, Portugal).

Techniques: Permeability, Sampling

Journal: Foods

Article Title: Effect of Coffee on the Bioavailability of Sterols

doi: 10.3390/foods11192935

Figure Lengend Snippet: Amount of LY transported and corresponding permeability coefficient in permeability assays through Caco-2 monolayers with a single time point assay of 6 h. Effect of increasing DHE concentrations (10, 50 and 100 µM) emulsified by GCA (10 mM), in the absence and presence of coffee extract D1, n = 2.

Article Snippet: Dulbecco’s modified Eagle’s medium high glucose (DMEM, 4.5 g/L glucose with 2 mM L-glutamine), sodium bicarbonate, nonessential amino acids (NEAA), penicillin (10,000 units/mL)–streptomycin (10 mg/mL) antibiotic, 0.25% ( w / v ) trypsin, ethylenediamine tetraacetic acid (EDTA), Hank’s balanced salt solution (HBSS), 4-2-hydroxyethyl-1-piperazineethanesulfonic acid (HEPES), Lucifer yellow CH di-potassium salt (LY), Hoechst 33,342, dehydroergosterol (DHE) and the sodium salts of glycocholic acid (GCA), glycodeoxycholic acid (GDCA) and glycochenodeoxycholic acid (GCDCA) were all purchased from Merck S.A (Algés, Portugal).

Techniques: Permeability

Journal: Foods

Article Title: Effect of Coffee on the Bioavailability of Sterols

doi: 10.3390/foods11192935

Figure Lengend Snippet: Confocal fluorescence images of Caco-2 monolayers stained for nuclei (blue) and ZO-1 (red). Representative optical section images along the z -axis (1 µm) are shown for cell monolayers after exposure during the permeability assay with: ( A ) GCA 10 mM, ( B ) DHE 50 µM + GCA 10 mM, ( C ) Coffee D1 + GCA 10 mM and ( D ) DHE 50 µM + coffee D1 + GCA 10 mM. The lower insets show the localization of the ZO-1 fluorescence in the orthogonal view of the z-x plane marked with a yellow line.

Article Snippet: Dulbecco’s modified Eagle’s medium high glucose (DMEM, 4.5 g/L glucose with 2 mM L-glutamine), sodium bicarbonate, nonessential amino acids (NEAA), penicillin (10,000 units/mL)–streptomycin (10 mg/mL) antibiotic, 0.25% ( w / v ) trypsin, ethylenediamine tetraacetic acid (EDTA), Hank’s balanced salt solution (HBSS), 4-2-hydroxyethyl-1-piperazineethanesulfonic acid (HEPES), Lucifer yellow CH di-potassium salt (LY), Hoechst 33,342, dehydroergosterol (DHE) and the sodium salts of glycocholic acid (GCA), glycodeoxycholic acid (GDCA) and glycochenodeoxycholic acid (GCDCA) were all purchased from Merck S.A (Algés, Portugal).

Techniques: Fluorescence, Staining, Permeability

Journal: Foods

Article Title: Effect of Coffee on the Bioavailability of Sterols

doi: 10.3390/foods11192935

Figure Lengend Snippet: Amount of DHE in suspension in the donor (apical) compartment in permeability assays using Caco-2 monolayers. The sterol DHE (10 μM) was emulsified by GCA (10 mM) in the absence (control) and in the presence of distinct coffee brews (expresso like coffee diluted 16 times). The amount of DHE is given as % of that in the sample before addition to the cells. Statistically significant differences from the control are indicated by * ( p < 0.05, n = 2).

Article Snippet: Dulbecco’s modified Eagle’s medium high glucose (DMEM, 4.5 g/L glucose with 2 mM L-glutamine), sodium bicarbonate, nonessential amino acids (NEAA), penicillin (10,000 units/mL)–streptomycin (10 mg/mL) antibiotic, 0.25% ( w / v ) trypsin, ethylenediamine tetraacetic acid (EDTA), Hank’s balanced salt solution (HBSS), 4-2-hydroxyethyl-1-piperazineethanesulfonic acid (HEPES), Lucifer yellow CH di-potassium salt (LY), Hoechst 33,342, dehydroergosterol (DHE) and the sodium salts of glycocholic acid (GCA), glycodeoxycholic acid (GDCA) and glycochenodeoxycholic acid (GCDCA) were all purchased from Merck S.A (Algés, Portugal).

Techniques: Permeability