Encyclopedia of Experiments: Neuroscience
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Transcript
Begin with a multi-well plate coated with a basement membrane matrix that includes extracellular polymers and adhesion proteins.
Seed the wells with a single-cell suspension of human embryonic stem cells in a growth medium containing a Rho-kinase inhibitor.
Incubate to allow stem cell attachment to the matrix.
Cells absorb the inhibitor, which blocks the Rho-kinase enzymes and prevents apoptotic cell death.
This promotes the cells' survival and maintains them in an undifferentiated state.
Replace the growth medium with a neural induction medium rich in growth factors and containing inhibitors that target specific ALK receptors.
These inhibitors interact with their receptors and block the BMP and TGF-β signaling pathways, preventing the cell differentiation into endodermal or mesodermal precursors.
This leads to selective differentiation of stem cells into ectodermal precursors.
These ectodermal precursors utilize nutrients and growth factors from the medium to multiply and later differentiate into neural progenitor cells.
