Generating Induced Pluripotent Stem Cell-Derived Cerebral Organoids
Transcription
Take colonies of induced pluripotent stem cells, or iPSCs, cultured on a supporting matrix.
Add a solution containing enzymes to dissociate the colonies into single cells.
Transfer the cells into a tube, centrifuge them, and discard the supernatant.
Resuspend the iPSCs in a growth medium.
Transfer the cells into a microplate containing biocompatible polymer fibers.
Centrifuge to force-aggregate iPSCs, forming embryoid bodies, or EBs, on the polymer scaffold.
Transition to a neural induction medium that promotes iPSC proliferation and differentiation into neural stem cells, progenitor cells, and immature neurons.
Upon incubation, the cells undergo self-organization.
Place the embryoid bodies onto a dimpled sheet and add drops containing extracellular matrix proteins.
Incubate to solidify the matrix, embedding the EBs.
Transfer the EBs into a microplate containing a neural maintenance medium. Incubate.
The EBs undergo cellular proliferation to form protrusions and buddings and differentiate into mature neurons, forming cerebral organoids.
