Generation of Neuronal Cells from Blood-Derived Pluripotent Stem Cells
Transkript
Place glass coverslips in 4-well plates and coat them with 1-to-5 diluted poly-L-ornithine in double-distilled water. After incubating the coverslips at 37 degrees Celsius for one hour, wash them with double-distilled water.
Slowly thaw 0.5 to 2 milligrams per milliliter of laminin, and add it to the top of coverslips. Incubate them at 37 degrees Celsius for two hours, then remove excess laminin by pipetting, and add neuronal medium N2 to the culture dishes.
Culture BD-derived CD45 negative cells in N2 medium on laminin-ornithine-coated glass coverslips for two days in an incubator, with 5% carbon dioxide at 37 degrees Celsius to initiate a neuronal differentiation of newly BD generated cells. Next, culture cells in neuronal differentiation medium, and place the plates in an incubator at 37 degrees Celsius and 5% carbon dioxide for 16 days.
