Generation of human cerebral organoids with a structured outer subventricular zone

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Research

Background:

• Outer radial glia (oRG) are crucial progenitor cells for neocortical expansion.
• Understanding oRG development is key to studying human brain development in vitro.
• Current methods lack efficient oRG generation in guided cortical organoids.

Purpose of the Study:

• To establish an in vitro differentiation method for generating oRG from human pluripotent stem cells (hPSCs).
• To investigate the role of STAT3 signaling and leukemia inhibitory factor (LIF) in oRG expansion.
• To enhance the cellular complexity of brain organoids for studying oRG.

Main Methods:

• Utilized human pluripotent stem cells (hPSCs) for cortical organoid differentiation.
• Activated the STAT3 signaling pathway using leukemia inhibitory factor (LIF).
• Incorporated neural crest-derived cortical pericytes to provide LIF in organoids.

Main Results:

• Developed a method that recapitulates the expansion of progenitor cells into the outer subventricular zone (oSVZ).
• Identified specific oRG markers (GFAP, LIFR, HOPX) in the generated oSVZ, matching fetal oRG.
• Demonstrated that incorporating LIF-producing pericytes mimics LIF treatment effects.

Conclusions:

• A novel cortical organoid differentiation method effectively generates oRG by activating STAT3 signaling.
• Increasing cellular complexity, specifically by adding LIF-producing pericytes, promotes oRG emergence.
• This platform facilitates routine study of oRG in hPSC-derived brain organoids, advancing research in neocortical development.