Method for promoting proliferation of neural stem cells
A neural stem cell technology, applied to nervous system cells, animal cells, vertebrate cells, etc., can solve the problems of single reagent and weak effect of cell proliferation
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Embodiment 1
[0033] Example 1 Different concentrations of VX-702 interfere with the proliferation of neural stem cells
[0034] The culture medium of the neural stem cell control group used basic culture medium: DMEM / F12 culture medium + 2% B27 (50×) + epidermal growth factor EGF (20ng / mL) + basic fibroblast growth factor bFGF (20ng / mL) + green / streptomycin (penicillin 100U / mL, streptomycin 0.1mg / mL) + amphotericin B (250ng / mL); the culture solution of the experimental group was added 1μM, 3μM, 5μM, 7μM, 10μM respectively on the basis of the control group VX-702.
[0035] Proceed as follows:
[0036] (1) After counting the neural stem cells cultured to the logarithmic growth phase, each well was divided into 3×10 3 Neural stem cells were seeded into 96-well plates, and each group was set up with 5 replicate wells.
[0037] (2) Divide the cells into blank wells (containing medium only), control wells (containing cells, medium, and no VX-702), and experimental wells (adding 1 μM, 3 μM, ...
Embodiment 2
[0041] Example 2 Detection of 7μM VX-702 Interfering with the Proliferation of Neural Stem Cells
[0042] The neural stem cells were divided into the control group and the experimental group. The culture medium of the control group was DMEM / F12 medium + 2% B27 (50×) + EGF (20ng / mL) + bFGF (20ng / mL) + penicillin / streptomycin ( Penicillin 100U / mL, streptomycin 0.1mg / mL) + amphotericin B (250ng / mL), the culture medium of the experimental group was added 7μM VX-702 on the basis of the control group.
[0043] Proceed as follows:
[0044](1) After the neural stem cells were in the logarithmic growth phase and counted on a counting plate, 1000 cells / well were planted in a 96-well plate, and each group was repeated for 5 wells, and 100 μL of ordinary medium was added to each well. Keep the cell volume in all wells equal, if the gap is too large, re-see the plate.
[0045] (2) In order to prevent the culture medium in the culture plate from being dried by the incubator, at the end of...
Embodiment 3
[0050] Example 3 7μM VX-702 Intervenes in the Formation of Neural Stem Cell Neurospheres
[0051] According to the method of steps (1) to (3) of Example 2, a control group and an experimental group were set up. The experimental group cultured neural stem cells with a medium added with 7 μM VX-702, and replaced the medium every 2 days for 3 days.
[0052] Such as image 3 As shown, after culturing for 3 days, observed under a phase-contrast microscope, the neural stem cell spheres in the experimental group gradually increased ( image 3 B).
[0053] The size and number of neural stem cell spheroids are as follows Figure 4 shown. Regardless of the neural stem cell spheres with a diameter of 20-50 μm, 51-80 μm or 81-110 μm, the number of the experimental group (VX-702) was far more than that of the control group (Ctrl), indicating that VX-702 (7 μM) can promote Proliferation of neural stem cells.
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