Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Generation of Spheroids from Human Pluripotent Stem Cells

Generation of Spheroids from Human Pluripotent Stem Cells

Transcription

Begin with colonies of human pluripotent stem cells or hPSCs attached to a suitable substrate in a culture medium.

Remove the medium. Add a neuronal medium containing specific inhibitors and incubate.

These inhibitors direct the maturation of hPSCs toward a neuronal lineage, forming neuroectodermal colonies. These are precursors to small, spherical structures called spheroids.

Remove the medium and wash with a salt buffer.

Incubate with a digestive enzyme to detach the colonies from the plate.

Add the neuronal medium to stop the enzymatic activity.

Collect the colonies and transfer them to a tube.

Allow them to settle, and then discard the supernatant.

Resuspend the colonies in a neuronal medium containing a growth factor.

Transfer the colonies to a multi-well plate and incubate.

This growth factor facilitates the self-organization and multiplication of neuroectodermal cells, leading them to form a spheroid.

Vidéos Connexes

Read Article