English
Generation of functional neurons from feeder-free, keratinocyte-derived equine induced pluripotent stem cells.
Abstract: Pluripotent stem cells (PSCs) offer unprecedented biomedical potential not only in relation to humans but also companion animals, particularly the horse. Despite this, attempts to generate bona fide equine embryonic stem cells have been unsuccessful. A very limited number of induced PSC lines have so far been generated from equine fibroblasts but their potential for directed differentiation into clinically relevant tissues has not been explored. In this study, we used retroviral vectors to generate induced pluripotent stem cells (iPSCs) with comparatively high efficiency from equine keratinocytes. Expression of endogenous PSC markers (OCT4, SOX2, LIN28, NANOG, DNMT3B, and REX1) was effectively restored in these cells, which could also form in vivo several tissue derivatives of the three germ layers, including functional neurons, keratinized epithelium, cartilage, bone, muscle, and respiratory and gastric epithelia. Comparative analysis of different reprogrammed cell lines revealed an association between the ability of iPSCs to form well-differentiated teratomas and the distinct endogenous expression of OCT4 and REX1 and reduced expression of viral transgenes. Importantly, unlike in previous studies, equine iPSCs were successfully expanded using simplified feeder-free culture conditions, constituting significant progress toward future biomedical applications. Further, under appropriate conditions equine iPSCs generated cells with features of cholinergic motor neurons including the ability to generate action potentials, providing the first report of functional cells derived from equine iPSCs. The ability to derive electrically active neurons in vitro from a large animal reveals highly conserved pathways of differentiation across species and opens the way for new and exciting applications in veterinary regenerative medicine.
Publication Date: 2014-03-25
PubMed ID: 24548115DOI: 10.1089/scd.2013.0565Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research focuses on generating induced pluripotent stem cells (iPSCs) from equine keratinocytes with high efficiency using retroviral vectors. These iPSCs can form several tissue derivatives, including functional neurons. The study signifies an important advancement in expanding iPSCs using feeder-free culture conditions, and details the creation of functional cells from equine iPSCs for the first time.
Generation of induced pluripotent stem cells (iPSCs)
- The researchers used retroviral vectors to generate iPSCs from equine keratinocytes (skin cells) more efficiently than previous methods.
- The resulting iPSCs effectively restored the expression of key pluripotent stem cell (cells that can become any type of cell) markers such as OCT4, SOX2, LIN28, NANOG, DNMT3B, and REX1.
Formation of diverse tissue derivatives
- The generated iPSCs were able to form several different types of tissues originating from the three germ layers, which are groups of cells in the embryo that develop into specific organ systems or tissues in the body.
- These tissues included functional neurons, keratinized epithelium, cartilage, bone, muscle, and respiratory and gastric epithelia.
Expanded Culture Conditions for iPSCs
- In contrast with previous studies, this research successfully expanded equine iPSCs in feeder-free culture conditions.
- This indicates considerable progress toward future applications of iPSCs in biomedical scenarios, reducing reliance on feeder layers that provide essential growth factors for stem cell survival and growth.
Derivation of Functional Neurons from Equine iPSCs
- For the first time, the study reports the generation of functional cells, particularly those with features of cholinergic motor neurons, from equine iPSCs.
- These neurons even exhibited the ability to generate action potentials, which are electrical signals crucial to neuron functioning.
- This achievement opens up new possibilities in veterinary regenerative medicine and further proves the conserved pathways of cell differentiation across different species.
Cite This Article
APA
Sharma R, Livesey MR, Wyllie DJ, Proudfoot C, Whitelaw CB, Hay DC, Donadeu FX.
(2014).
Generation of functional neurons from feeder-free, keratinocyte-derived equine induced pluripotent stem cells.
Stem Cells Dev, 23(13), 1524-1534.
https://doi.org/10.1089/scd.2013.0565
Publication
Researcher Affiliations
- 1 The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh , Midlothian, United Kingdom .
MeSH Terms
- Animals
- Biomarkers / metabolism
- Cell Culture Techniques
- Cell Differentiation
- Feeder Cells
- Gene Expression
- Horses
- Induced Pluripotent Stem Cells / physiology
- Keratinocytes / physiology
- Mice, Inbred NOD
- Mice, SCID
- Neurons / physiology
- Spheroids, Cellular / cytology
- Transcription Factors / genetics
- Transcription Factors / metabolism
Grant Funding
- BBS/E/D/05251442 / Biotechnology and Biological Sciences Research Council
- BBS/E/D/05251443 / Biotechnology and Biological Sciences Research Council
- BBS/E/D/05251444 / Biotechnology and Biological Sciences Research Council
- BBS/E/D/05251445 / Biotechnology and Biological Sciences Research Council
Citations
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