siRNA-Mediated Gene Silencing in Neural Stem Cells
Transcrição
To perform siRNA knockdown in O9-1 cells recover and seed the cells. Dilute liposomes in an appropriate volume of serum-free media. Then, dilute siRNA in serum-free media according to the text protocol. After this, add the diluted siRNA to the diluted liposomes according to the manufacturer's instructions. Mix the solution by pipetting and incubate for 5 minutes at room temperature.
Next, add an appropriate volume of siRNA-lipid complex to the cells. Then, incubate the cells for 24 hours in a standard cell culture incubator. O9-1 cells can differentiate into different cell types under specific differentiation culture conditions. To differentiate O9-1 cells into osteoblasts, prepare osteogenic differentiation media according to the text protocol. Finally, detect the osteoblast marker osteocalcin in differentiated osteoblasts by immunostaining.
