Generation of Functional Neurons from Feeder-Free, Keratinocyte-Derived Equine Induced Pluripotent Stem Cells
| Autor: | David C. Hay, Christopher Proudfoot, Christopher Bruce Alexander Whitelaw, Matthew R. Livesey, Ruchi Sharma, Francesc X Donadeu, David J. A. Wyllie |
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| Rok vydání: | 2014 |
| Předmět: |
Keratinocytes
Homeobox protein NANOG KOSR Cellular differentiation Induced Pluripotent Stem Cells Cell Culture Techniques Gene Expression Mice SCID Biology Directed differentiation SOX2 Mice Inbred NOD Spheroids Cellular Animals Horses Induced pluripotent stem cell Neurons Feeder Cells Cell Differentiation Cell Biology Hematology Embryonic stem cell Cell biology Immunology Stem cell Biomarkers Transcription Factors Developmental Biology |
| Zdroj: | Stem Cells and Development. 23:1524-1534 |
| ISSN: | 1557-8534 1547-3287 |
| DOI: | 10.1089/scd.2013.0565 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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