Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids
- University of Cambridge, United Kingdom
- University College London, United Kingdom
Abstract
The embryonic mouse lung is a widely used substitute for human lung development. For example, attempts to differentiate human pluripotent stem cells to lung epithelium rely on passing through progenitor states that have only been described in mouse. The tip epithelium of the branching mouse lung is a multipotent progenitor pool that self-renews and produces differentiating descendants. We hypothesized that the human distal tip epithelium is an analogous progenitor population and tested this by examining morphology, gene expression and in vitro self-renewal and differentiation capacity of human tips. These experiments confirm that human and mouse tips are analogous and identify signalling pathways that are sufficient for long-term self-renewal of human tips as differentiation-competent organoids. Moreover, we identify mouse-human differences, including markers that define progenitor states and signalling requirements for long-term self-renewal. Our organoid system provides a genetically-tractable tool that will allow these human-specific features of lung development to be investigated.
Data availability
-
Expression data from fresh human embryonic lung epithelial tip and stalk cells and cultured organoids derived from tip and stalkPublicly available at the NCBI Gene Expression Omnibus (accession no: GSE95860).
-
Expression data from mouse embryonic lung epithelial tip progenitor cellsPublicly available at the NCBI Gene Expression Omnibus (accession no: GSM1968996).
-
Microdissected E11.5 lung epithelial tip biological replicate 2Publicly available at the NCBI Gene Expression Omnibus (accession no: GSM1968997).
-
Microdissected E11.5 lung epithelial tip biological replicate 3Publicly available at the NCBI Gene Expression Omnibus (accession no: GSM1968998).
-
Microdissected E11.5 lung epithelial tip biological replicate 4Publicly available at the NCBI Gene Expression Omnibus (accession no: GSM1968999).
-
Microdissected E11.5 lung epithelial tip biological replicate 5Publicly available at the NCBI Gene Expression Omnibus (accession no: GSM1969000).
Article and author information
Author details
Funding
Medical Research Council (G0900424)
- Emma L Rawlins
Wellcome (PhD programme for clinicians fellowship)
- Marko Z Nikolić
Addenbrooke's Charitable Trust, Cambridge University Hospitals (N/A)
- Marko Z Nikolić
Ikerbasque, Basque Foundation for Science (N/A)
- Usua Laresgoiti
Cancer Research UK (C6946/A14492)
- Emma L Rawlins
Medical Research Council (MR/P009581/1)
- Emma L Rawlins
Wellcome (Clinical PhD Fellowship)
- Jo-Anne Johnson
Wellcome (92096)
- Emma L Rawlins
Fundacio Universitaria Agusti Pedro i Pons (Short term fellowship)
- Oriol Caritg
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This research has been regulated under the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012 following ethical review by the University of Cambridge and University College London Animal Welfare and Ethical Review Body (AWERB). All experiments were thereafter conducted according to Home Office project licenses PPL 70/8012 (Emma Rawlins, University of Cambridge) and 70/7607 (Adam Giangreco, UCL).
Human subjects: Human embryonic and foetal lung tissue. Human embryonic and foetal lungs were obtained from terminations of pregnancy from Cambridge University Hospitals NHS Foundation Trust under permission from NHS Research Ethical Committee (96/085) and the Joint MRC/Wellcome Trust Human Developmental Biology Resource (London and Newcastle, grant 099175/Z/12/Z, www.hdbr.org). Informed consent and consent to publish was obtained.Human adult lung tissue. Fresh healthy adult lung tissue (background tissue from lobectomies for lung cancer) was obtained from Papworth Hospital NHS Foundation Trust (Research Tissue Bank Generic REC approval, Tissue Bank Project number T01939). Informed consent and consent to publish was obtained.
Copyright
© 2017, Nikolić 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.
Metrics
-
- 10,326
- views
-
- 1,690
- downloads
-
- 239
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids
eLife 6:e26575.
https://doi.org/10.7554/eLife.26575
Further reading
-
- Stem Cells and Regenerative Medicine
A new source of progenitor cells can now be used to study hidden aspects of human lung development and pediatric lung disease.
-
- Genetics and Genomics
- Stem Cells and Regenerative Medicine
Retinal degeneration in mammals causes permanent loss of vision, due to an inability to regenerate naturally. Some non-mammalian vertebrates show robust regeneration, via Muller glia (MG). We have recently made significant progress in stimulating adult mouse MG to regenerate functional neurons by transgenic expression of the proneural transcription factor Ascl1. While these results showed that MG can serve as an endogenous source of neuronal replacement, the efficacy of this process is limited. With the goal of improving this in mammals, we designed a small molecule screen using sci-Plex, a method to multiplex up to thousands of single-nucleus RNA-seq conditions into a single experiment. We used this technology to screen a library of 92 compounds, identified, and validated two that promote neurogenesis in vivo. Our results demonstrate that high-throughput single-cell molecular profiling can substantially improve the discovery process for molecules and pathways that can stimulate neural regeneration and further demonstrate the potential for this approach to restore vision in patients with retinal disease.
