Development of human nervous tissue upon differentiation of embryonic stem cells in three-dimensional culture

Abstract

Researches on neural differentiation using embryonic stem cells (ESC) require analysis of neurogenesis in conditions mimicking physiological cellular interactions as closely as possible. In this study, we report an air-liquid interface-based culture of human ESC. This culture system allows three-dimensional cell expansion and neural differentiation in the absence of added growth factors. Over a 3-month period, a macroscopically visible, compact tissue developed. Histological coloration revealed a dense neural-like neural tissue including immature tubular structures. Electron microscopy, immunochemistry, and electrophysiological recordings demonstrated a dense network of neurons, astrocytes, and oligodendrocytes able to propagate signals. Within this tissue, tubular structures were niches of cells resembling germinal layers of human fetal brain. Indeed, the tissue contained abundant proliferating cells expressing markers of neural progenitors. Finally, the capacity to generate neural tissues on air-liquid interface differed for different ESC lines, confirming variations of their neurogenic potential. In conclusion, this study demonstrates in vitro engineering of a human neural-like tissue with an organization that bears resemblance to early developing brain. As opposed to previously described methods, this differentiation (a) allows three-dimensional organization, (b) yields dense interconnected neural tissue with structurally and functionally distinct areas, and (c) is spontaneously guided by endogenous developmental cues.

Engineering of Midbrain Organoids Containing Long-Lived Dopaminergic Neurons

“…The alternative to presynthesized scaffolds is the generation of an extracellular matrix by the cells themselves in 3D tissue culture. Previous studies from our laboratory have shown that this is basically possible [13,14].…”

Monocrotophos in Gandaman village: India school lunch deaths and need for improved toxicity testing

“…Thanks to induced PSC technology (Takahashi and Yamanaka 2006), the ethnic diversity of humans can be included into the analysis through a careful selection of PSC lines (Fakunle and Loring 2012). Three-dimensional culture techniques (Hoelting et al 2013) have enabled the generation of PSC-derived organoids (e.g., Preynat-Seauve et al 2009), allowing putative toxic compounds to be tested on tissues rather than on isolated cells. Finally, the use of PSCs to generate mice with humanized organs (e.g., Takebe et al 2013) provides new opportunities to represent physiological complexity.…”

Antitumor Compound Testing in Glioblastoma Organotypic Brain Cultures

“…recently, a method that allows brain embryonic stem cell expansion has been described using an air-liquid interface-based method. 12 We show here that the use of such an air-liquid interface system to co-cultivate brain cells together with glioblastoma cells recapitulates features of clinical Gbm and offers a unique simple tool to assess the pharmacodynamic properties of potential anticancer drugs.…”