Self-Organized Formation of Polarized Cortical Tissues from ESCs and Its Active Manipulation by Extrinsic Signals

Abstract

Here, we demonstrate self-organized formation of apico-basally polarized cortical tissues from ESCs using an efficient three-dimensional aggregation culture (SFEBq culture). The generated cortical neurons are functional, transplantable, and capable of forming proper long-range connections in vivo and in vitro. The regional identity of the generated pallial tissues can be selectively controlled (into olfactory bulb, rostral and caudal cortices, hem, and choroid plexus) by secreted patterning factors such as Fgf, Wnt, and BMP. In addition, the in vivo-mimicking birth order of distinct cortical neurons permits the selective generation of particular layer-specific neurons by timed induction of cell-cycle exit. Importantly, cortical tissues generated from mouse and human ESCs form a self-organized structure that includes four distinct zones (ventricular, early and late cortical-plate, and Cajal-Retzius cell zones) along the apico-basal direction. Thus, spatial and temporal aspects of early corticogenesis are recapitulated and can be manipulated in this ESC culture.

Sip1 regulates sequential fate decisions by feedback signaling from postmitotic neurons to progenitors

“…An internal clock-like mechanism capable of counting the number of cell divisions that a progenitor cell goes through has also been proposed. Recently, it was shown that embryonic stem cells can be driven toward cells with dorsal telencephalic fate by blocking Shh-mediated signaling 30,31 . However, the amount of upper-layer neurons generated in these cultures was lower, suggesting that extrinsic factors that influence upper-layer cell fate were present.…”

Novel and Robust Transplantation Reveals the Acquisition of Polarized Processes by Cortical Cells Derived from Mouse and Human Pluripotent Stem Cells

“…Indeed, the generation of the different neuronal subtypes found within the six distinct layers of the mammalian cortex from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) has been established in adherent monolayer culture systems [1][2][3] and self-organized cultures with polarized cytoarchitecture [4]. With these types of approaches, the entire neurogenic process, from undifferentiated stem cells to terminally differentiated neurons, can be tracked in the defined conditions of a culture dish.…”

Translational potential of human brain organoids

“…Independently Yoshiki Sasai's group explored directed differentiation of hPSCs after embryoid body (EB) formation, and developed a method known as the serum‐free culture of embryoid bodies after quick reaggregation (SEEBq) 25, 26. When specific neural inductive molecules were added, they generated polarized neural tissues 25, 26…”