Generating Cortical Pyramidal Neurons from Human Neural Stem Cells

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.