A Technique for Generating Neural Progenitor Cells from Mouse Embryonic Stem Cells
Transcrição
Add 1.5 million mouse embryonic stem cells into a non-adhesive bacterial dish in 10 milliliters of basal differentiation medium, and incubate the plate at 37 degrees Celsius and 5% carbon dioxide to allow for embryoid body formation.
After two days, transfer the cell aggregates into 15-milliliter centrifuge tubes, and let them settle by gravity. Remove the supernatant and add 10 milliliters of fresh basal differentiation medium to resuspend the embryoid bodies. Then, replant them into a new non-adhesive dish and incubate them for another two days.
Collect and seed about 50 embryoid bodies in 2 milliliters of basal differentiation medium per well onto a gelatin-coated six-well plate. For retinoic acid induction, add 2 microliters of RHA stock into each well to a final concentration of 1 micromolar. Return the plate to the incubator and differentiate the cells for another four days.
