An In Vitro Method to Generate Human Brain Organoids
Transcription
Begin with a culture of neurospheres, free-floating clusters of neuroepithelial progenitor cells.
Transfer these neurospheres onto an embedding sheet.
Remove media. Add a drop of extracellular membrane matrix containing proteins and growth factors to each neurosphere.
Incubate for the matrix to solidify, embedding the neurospheres within it.
Now, transfer the matrix-embedded neurospheres using neurosphere media to a culture plate containing neurosphere media.
Incubate. The extracellular membrane matrix and its constituents facilitate neuroepithelial progenitor cell differentiation into neuroepithelial cells which proliferate and form fluid-filled lumens.
Post-incubation, transfer neurospheres into a spinner flask containing pre-warmed brain organoid media. Incubate with stirring for enhanced oxygen and nutrient diffusion, promoting cellular growth.
Growth factors and small molecule inhibitors in media promote neural differentiation and develop into cells of specific brain regions, forming mature brain organoids.
