Generating Induced Pluripotent Stem Cell-Derived Cerebral Organoids

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.

Aspirate the medium with a pipette, and wash the cells once with DPBS. Then, add 500 microliters of cell detachment solution, or 0.5 millimolar EDT