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.
Reviewing Editor
- Marianne E Bronner, California Institute of Technology, United States
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).
Version history
- Received: February 5, 2021
- Preprint posted: February 8, 2021 (view preprint)
- Accepted: February 2, 2022
- Accepted Manuscript published: February 21, 2022 (version 1)
- Version of Record published: March 17, 2022 (version 2)
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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Large-scale cell flow characterizes gastrulation in animal development. In amniote gastrulation, particularly in avian gastrula, a bilateral vortex-like counter-rotating cell flow, called ‘polonaise movements’, appears along the midline. Here, through experimental manipulations, we addressed relationships between the polonaise movements and morphogenesis of the primitive streak, the earliest midline structure in amniotes. Suppression of the Wnt/planar cell polarity (PCP) signaling pathway maintains the polonaise movements along a deformed primitive streak. Mitotic arrest leads to diminished extension and development of the primitive streak and maintains the early phase of the polonaise movements. Ectopically induced Vg1, an axis-inducing morphogen, generates the polonaise movements, aligned to the induced midline, but disturbs the stereotypical cell flow pattern at the authentic midline. Despite the altered cell flow, induction and extension of the primitive streak are preserved along both authentic and induced midlines. Finally, we show that ectopic axis-inducing morphogen, Vg1, is capable of initiating the polonaise movements without concomitant PS extension under mitotic arrest conditions. These results are consistent with a model wherein primitive streak morphogenesis is required for the maintenance of the polonaise movements, but the polonaise movements are not necessarily responsible for primitive streak morphogenesis. Our data describe a previously undefined relationship between the large-scale cell flow and midline morphogenesis in gastrulation.
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Geometric criteria can be used to assess whether cell intercalation is active or passive during the convergent extension of tissue.
