Mouse embryonic stem cells can differentiate via multiple paths to the same state
- Harvard Medical School, United States
Abstract
In embryonic development, cells differentiate through stereotypical sequences of intermediate states to generate particular mature fates. By contrast, driving differentiation by ectopically expressing terminal transcription factors (direct programming) can generate similar fates by alternative routes. How differentiation in direct programming relates to embryonic differentiation is unclear. We applied single-cell RNA sequencing to compare two motor neuron differentiation protocols: a standard protocol approximating the embryonic lineage, and a direct programming method. Both initially undergo similar early neural commitment. Later, the direct programming path diverges into a novel transitional state rather than following the expected embryonic spinal intermediates. The novel state in direct programming has specific and uncharacteristic gene expression. It forms a loop in gene expression space that converges separately onto the same final motor neuron state as the standard path. Despite their different developmental histories, motor neurons from both protocols structurally, functionally, and transcriptionally resemble motor neurons isolated from embryos.
Data availability
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Single cell RNA-seq data for: Mouse embryonic stem cells can differentiate via multiple paths to the same statePrivate pre-publication access available at the NCBI Gene Expression Omnibus (accession no: GSE97391).
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Microarray data for: Mouse embryonic stem cells can differentiate via multiple paths to the same statePrivate pre-publication access available at the NCBI Gene Expression Omnibus (accession no: GSE97391).
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Droplet barcoding for single cell transcriptomics applied to embryonic stem cellsPublicly available at the NCBI Gene Expression Omnibus (accession no: GSE65525).
Article and author information
Author details
Funding
National Institutes of Health (R21 HD087723)
- Victor C Li
- Marc W Kirschner
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Martin Pera, University of Melbourne, Australia
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#16-01-3080R) of Boston Children's Hospital, and (#IS00000137, #1648, #1648a, #1648b) of Harvard Medical School. The Hb9-GFP embryos were collected from a pregnant mouse after euthanasia by inhalation of CO2, and every effort was made to minimize suffering.
Version history
- Received: March 17, 2017
- Accepted: October 5, 2017
- Accepted Manuscript published: October 9, 2017 (version 1)
- Version of Record published: October 19, 2017 (version 2)
- Version of Record updated: December 21, 2017 (version 3)
Copyright
© 2017, Briggs 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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Mouse embryonic stem cells can differentiate via multiple paths to the same state
eLife 6:e26945.
https://doi.org/10.7554/eLife.26945
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