A Co-culture Method to Model Neuron-Oligodendrocyte Interactions
Trascrizione
Take human-induced neurons, or iNs, cultured on an extracellular matrix.
Remove the media and add digestive enzymes that disrupt the matrix, dissociating the cells.
Transfer the iNs into a tube. Centrifuge the suspension and discard the supernatant containing the enzymes.
Resuspend the iNs in a co-culture medium; transfer them onto a culture of induced oligodendrocyte precursor cells, or iOPCs.
iOPCs are precursors to oligodendrocytes that form myelin sheaths around the axons of neurons.
Upon incubation, the iOPCs form synaptic connections with the iNs. Trophic factors released by the iOPCs and growth factors in the medium promote iN maturation into neurons, exhibiting increased cellular processes that form synaptic connections with other neurons.
The electrical activity of the mature neurons and growth factors in the medium induces iOPC maturation into oligodendrocytes.
The oligodendrocytes extend cellular processes, forming myelin sheaths around the neuronal axons.
The established co-culture is ready for analysis.
