Generation of human cerebral organoids with a structured outer subventricular zone.

Outer radial glia (oRG) emerge as cortical progenitor cells that support the development of an enlarged outer subventricular zone (oSVZ) and the expansion of the neocortex. The in vitro generation of oRG is essential to investigate the underlying mechanisms of human neocortical development and expansion. By activating the STAT3 signaling pathway using leukemia inhibitory factor (LIF), which is not expressed in guided cortical organoids, we define a cortical organoid differentiation method from human pluripotent stem cells (hPSCs) that recapitulates the expansion of a progenitor pool into the oSVZ. The oSVZ comprises progenitor cells expressing specific oRG markers such as GFAP, LIFR, and HOPX, closely matching human fetal oRG. Finally, incorporating neural crest-derived LIF-producing cortical pericytes into cortical organoids recapitulates the effects of LIF treatment. These data indicate that increasing the cellular complexity of the organoid microenvironment promotes the emergence of oRG and supports a platform to study oRG in hPSC-derived brain organoids routinely.

Walsh RM ，Luongo R ，Giacomelli E ，Ciceri G ，Rittenhouse C ，Verrillo A ，Galimberti M ，Bocchi VD ，Wu Y ，Xu N ，Mosole S ，Muller J ，Vezzoli E ，Jungverdorben J ，Zhou T ，Barker RA ，Cattaneo E ，Studer L ，Baggiolini A ... -  《Cell Reports》

Generation of human cerebral organoids with a structured outer subventricular zone.

Walsh R ，Giacomelli E ，Ciceri G ，Rittenhouse C ，Galimberti M ，Wu Y ，Muller J ，Vezzoli E ，Jungverdorben J ，Zhou T ，Barker RA ，Cattaneo E ，Studer L ，Baggiolini A ... -  《-》

Protocol for generating human cortical organoids enriched in outer radial glia by guided differentiation.

The generation of human pluripotent stem cell (hPSC)-derived brain organoids is continuously refined, enhancing their reproducibility and complexity. Here, we present a guided differentiation protocol for generating cortical forebrain organoids and cortico-pericyte (CP)assembloids composed of a robust outer radial glia (oRG) population and an expanded outer subventricular zone (oSVZ). We describe the steps to generate hPSC-derived cortical organoids (COs), cortical pericytes, and CP assembloids. Moreover, we outline the procedures to characterize the organoids by immunostaining and to perform single-cell dissociation. For complete details on the use and execution of this protocol, please refer to Walsh et al.1.

Luongo R ，Walsh RM ，Verrillo A ，Studer L ，Baggiolini A ... -  《-》

Recapitulating cortical development with organoid culture in vitro and modeling abnormal spindle-like (ASPM related primary) microcephaly disease.

Li R ，Sun L ，Fang A ，Li P ，Wu Q ，Wang X ... -  《-》

Neurogenic radial glia in the outer subventricular zone of human neocortex.

Hansen DV ，Lui JH ，Parker PR ，Kriegstein AR ... -  《-》