Modeling neural tube development by differentiation of human embryonic stem cells in a microfluidic WNT gradient.
Author(s): Pedro Rifes, Marc Isaksson, Gaurav Singh Rathore, Patrick Aldrin-Kirk, Oliver Knights Møller, Guido Barzaghi, Julie Lee, Kristoffer Lihme Egerod, Dylan M. Rausch, Malin Parmar, Tune H. Pers, Thomas Laurell & Agnete Kirkeby
The Novo Nordisk Foundation Center for Stem Cell Biology (DanStem)06/22/2020
The study of brain development in humans is limited by the lack of tissue samples and suitable in vitro models. Here, we model early human neural tube development using human embryonic stem cells cultured in a microfluidic device. The approach, named microfluidic-controlled stem cell regionalization (MiSTR), exposes pluripotent stem cells to signaling gradients that mimic developmental patterning. Using a WNT-activating gradient, we generated a neural tissue exhibiting progressive caudalization from forebrain to midbrain to hindbrain, including formation of isthmic organizer characteristics. Single-cell transcriptomics revealed that rostro-caudal organization was already established at 24 h of differentiation, and that the first markers of a neural-specific transcription program emerged in the rostral cells at 48 h. The transcriptomic hallmarks of rostro-caudal organization recapitulated gene expression patterns of the early rostro-caudal neural plate in mouse embryos. Thus, MiSTR will facilitate research on the factors and processes underlying rostro-caudal neural tube patterning.
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