Human spinal cord in vitro differentiation pace is initially maintained in heterologous embryonic environments
- University of Dundee, United Kingdom
Abstract
Species-specific differentiation pace in-vitro indicates that some aspects of neural differentiation are governed by cell intrinsic properties. Here we describe a novel in-vitro human neural-rosette assay that recapitulates dorsal spinal cord differentiation but proceeds more rapidly than in the human embryo, suggesting that it lacks endogenous signalling dynamics. To test whether in-vitro conditions represent an intrinsic differentiation pace, human iPSC-derived neural rosettes were challenged by grafting into the faster differentiating chicken embryonic neural tube iso-chronically, or hetero-chronically into older embryos. In both contexts in-vitro differentiation pace was initially unchanged, while long-term analysis revealed iso-chronic slowed and hetero-chronic conditions promoted human neural differentiation. Moreover hetero-chronic conditions did not alter the human neural differentiation programme, which progressed to neurogenesis, while the host embryo advanced into gliogenesis. This study demonstrates that intrinsic properties limit human differentiation pace, and that timely extrinsic signals are required for progression through an intrinsic human neural differentiation programme.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data (all numerical meta data) are provided as excel tables aligned to the relevant figure.
Article and author information
Author details
Funding
Wellcome Trust (WT102817AIA)
- Kate G Storey
Wellcome Trust (WT102817/Z/13/A)
- Kate G Storey
Wellcome Trust (WT101468)
- Kate G Storey
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Human subjects: Human embryonic tissue (4 to 6 weeks of gestation) was obtained from the MRC/ Wellcome-Trust (grant no. 006237/1) funded Human Developmental Biology Resource (HDBR; www.hdbr.org) with appropriate maternal written consent and approval from the London Fulham Research Ethics Committee (18/LO/ 0822) and the Newcastle and North Tyneside NHS Health Authority Joint Ethics Committee (08/H0906/21+5). HDBR is regulated by the UK Human Tissue Authority (HTA; www.hta.gov.uk) and operates in accordance with the relevant HTA codes of practice. This work was part of project no. 200407 registered with the HDBR.Human ESC lines H9 and H1 expressing DCXCit/Y were provided by WiCel and all work with hESCs was undertaken in approval of the UK Stem Cell Bank steering committee (license number SCSC14-29).
Copyright
© 2022, Dady et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Human spinal cord in vitro differentiation pace is initially maintained in heterologous embryonic environments
eLife 11:e67283.
https://doi.org/10.7554/eLife.67283
Further reading
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- Developmental Biology
During the first lineage segregation, mammalian embryos generate the inner cell mass (ICM) and trophectoderm (TE). ICM gives rise to the epiblast (EPI) that forms all cell types of the body, an ability referred to as pluripotency. The molecular mechanisms that induce pluripotency in embryos remain incompletely elucidated. Using knockout (KO) mouse models in conjunction with low-input ATAC-seq and RNA-seq, we found that Oct4 and Sox2 gradually come into play in the early ICM, coinciding with the initiation of Sox2 expression. Oct4 and Sox2 activate the pluripotency-related genes through the putative OCT-SOX enhancers in the early ICM. Furthermore, we observed a substantial reorganization of chromatin landscape and transcriptome from the morula to the early ICM stages, which was partially driven by Oct4 and Sox2, highlighting their pivotal role in promoting the developmental trajectory toward the ICM. Our study provides new insights into the establishment of the pluripotency network in mouse preimplantation embryos.
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