Establishment method of inflammation model of mice primary colonic epithelial cells induced by dextran sodium sulfate combined with lipopolysaccharides
A technology of dextran sodium sulfate and establishment method, which is applied in the field of establishment of mouse primary colonic mucosal epithelial cell inflammation model, can solve the problem of low cost, and achieve the effect of low cost and easy operation
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Embodiment 1
[0041] Effects of different concentrations of dextran sodium sulfate treatment on cell viability:
[0042] (1) Experimental grouping and processing
[0043] The cell control group and the dextran sodium sulfate group with different concentration gradients (0.31%, 0.62%, 1.25%, 2.5%, 5%, 10%, 12.5%, 15% w / v) were respectively set up, with 2% (w / v) FBS iCell primary epithelial cell culture medium (product number: PriMed-iCELL-001) to adjust the concentration of mouse primary colonic mucosal epithelial cells to 1×10 5 cells / mL, and seeded in 96-well plate, 100 μL per well, 37°C, 5% CO 2 Cultivate overnight in the incubator; add 100 μL of different concentrations of dextran sodium sulfate diluted with serum-free iCell primary epithelial cell culture medium to each well of the experimental group, and add the same amount of serum-free iCell primary epithelial cell culture medium to the blank control group Cell culture medium, 37°C, 5% CO 2 Continue to grow in the incubator.
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Embodiment 2
[0047] Effects of treatment with different concentrations of lipopolysaccharide combined with 10% dextran sodium sulfate on the levels of TNF-α and IL-1β in cell culture supernatant
[0048] (1) Experimental grouping and processing
[0049] Cell control group and lipopolysaccharide group with different concentration gradients (1 μg / mL, 10 μg / mL, 100 μg / mL) were respectively set up, adjusted with iCell primary epithelial cell culture medium (product number: PriMed-iCELL-001) containing 2% FBS. The concentration of mouse primary colonic mucosal epithelial cells was 1×10 5 cells / mL, seeded in 24-well plate, 1000 μL per well, 37°C, 5% CO 2 Cultivate overnight in the incubator; add 1000 μL of 10% dextran sodium sulfate diluted with serum-free iCell primary epithelial cell culture medium to the experimental group, at 37°C, 5% CO 2 After being stimulated in the incubator for 3 hours, suck out the supernatant, add 100 μL PBS to each well and soak for 10 seconds, gently suck out and ...
Embodiment 3
[0053] 1. The experimental data were analyzed by one-way analysis of variance (One-Way ANOVA) using SSP20.0 statistical software, and compared between Duncan groups.
[0054] 2. Experimental results
[0055] (1) Effects of different concentrations of dextran sodium sulfate on the inhibitory rate of primary colonic mucosal epithelial cells in mice
[0056] The result is as figure 1 As shown, as the concentration of dextran sodium sulfate increased, the inhibition rate gradually increased; and as the action time increased, the cell inhibition rate gradually increased. Among them, 10% dextran sodium sulfate treated primary colonic epithelial cells of mice for 2 hours and 3 hours can make the inhibition rate of the cells reach 50% (P<0.05).
[0057] (2) The effect of 10% dextran sodium sulfate combined with lipid treatment on the TNF-α content of primary colonic mucosal epithelial cells of mice
[0058] The result is as figure 2 As shown, in the three stimulation times of 3h,...
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