Generating Cortical Pyramidal Neurons from Human Neural Stem Cells
Transcrição
In a culture plate, coat a coverslip with polyornithine, a positively charged polymer, and incubate.
Polyornithine forms a mesh-like layer that helps negatively charged cells stick to the coverslip.
Remove the solution and wash the coverslips with a buffered salt solution to remove unbound polyornithine.
Add a solution containing an extracellular matrix protein called laminin and incubate.
Laminin forms a matrix over the polyornithine-coated coverslip, enhancing cell attachment.
Discard the solution and take a small number of neural stem cells suspended in a growth factor-free medium.
Add this low-density culture over the prepared coverslip in a slow rotating movement to evenly distribute the cells.
Initially, growth factor deprivation promotes cell division.
Over time, stem cells differentiate into neurons with long projections
Eventually, neurons produce spine-like structures over their projections, like the cortical pyramidal neurons.
