Generation of human cerebral organoids with a structured outer subventricular zone

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Biology

Background:

• Mammalian outer radial glia (oRG) are crucial cortical progenitor cells driving neocortical expansion.
• Generating oRG in vitro is vital for modeling human neocortical development and expansion.
• Existing cerebral organoid models lack LIF, a key factor for oRG development.

Approach:

• Developed a cerebral organoid differentiation method using human pluripotent stem cells (hPSCs).
• Activated the STAT3 pathway with LIF to promote oRG generation and recapitulate outer subventricular zone (oSVZ) expansion.
• Incorporated hPSC-derived brain vascular pericytes to mimic endogenous LIF production.

Key Points:

• The method successfully generated a structured oSVZ with oRG markers (GFAP, LIFR, HOPX), mirroring in vivo human oRG.
• Cortical neurons in this microenvironment exhibited accelerated maturation, enhanced metabolic, and functional activity.
• LIF-producing pericytes in organoids mimicked LIF treatment effects, promoting oRG appearance.

Conclusions:

• Activating the STAT3 pathway is key to generating oRG in hPSC-derived cerebral organoids.
• The cellular complexity of the cortical microenvironment, including vascular cells, supports oRG development.
• This platform enables routine study of oRG in hPSC-derived brain organoids.