Establishing a Müller Glial Cell Culture Obtained from Mouse Retinas
Transcrição
Take mouse retinas containing Müller Glial, or MG cells, and neurons.
Place them in a dissociation solution with digestive enzymes and incubate with gentle rocking to ensure even enzyme distribution.
These enzymes break down the extracellular matrix, detaching individual cells.
Pipette the retinas up and down several times to release the cells.
Add an inhibitor to halt enzyme activity, then centrifuge.
Discard the supernatant.
Resuspend the cells in media containing a growth factor specific to MG cells.
Transfer the cells to a well and incubate.
Over time, the growth factor in the media facilitates the growth and multiplication of MG cells while the non-dividing neuronal cells perish.
Remove the media. Wash with a salt buffer.
Incubate with a digestive enzyme to detach the cells from the well.
Collect the cells in a tube and then centrifuge.
Discard the supernatant containing the dead neuronal cells.
Resuspend the MG cells in media for further analysis.
