Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer
Induced pluripotent stem cells (iPSCs) share similar characteristics of indefinite in vitro growth with embryonic stem cells (ESCs) and may therefore serve as a useful tool for the targeted genetic modification of farm animals via nuclear transfer (NT). Derivation of stable ESC lines from farm anima...
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| Format: | Article |
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Mary Ann Liebert
2015
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| Online Access: | https://eprints.nottingham.ac.uk/28725/ |
| _version_ | 1848793631294488576 |
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| author | German, Sergio D. Campbell, Keith H.S. Thornton, Elisabeth McLachlan, Gerry Sweetman, Dylan Alberio, Ramiro |
| author_facet | German, Sergio D. Campbell, Keith H.S. Thornton, Elisabeth McLachlan, Gerry Sweetman, Dylan Alberio, Ramiro |
| author_sort | German, Sergio D. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Induced pluripotent stem cells (iPSCs) share similar characteristics of indefinite in vitro growth with embryonic stem cells (ESCs) and may therefore serve as a useful tool for the targeted genetic modification of farm animals via nuclear transfer (NT). Derivation of stable ESC lines from farm animals has not been possible, therefore, it is important to determine whether iPSCs can be used as substitutes for ESCs in generating genetically modified cloned farm animals. We generated ovine iPSCs by conventional retroviral transduction using the four Yamanaka factors. These cells were basic fibroblast growth factor (bFGF)- and activin A-dependent, showed persistent expression of the transgenes, acquired chromosomal abnormalities, and failed to activate endogenous NANOG. Nonetheless, iPSCs could differentiate into the three somatic germ layers in vitro. Because cloning of farm animals is best achieved with diploid cells (G1/G0), we synchronized the iPSCs in G1 prior to NT. Despite the cell cycle synchronization, preimplantation development of iPSC-NT embryos was lower than with somatic cells (2% vs. 10% blastocysts, p<0.01). Furthermore, analysis of the blastocysts produced demonstrated persistent expression of the transgenes, aberrant expression of endogenous SOX2, and a failure to activate NANOG consistently. In contrast, gene expression in blastocysts produced with the parental fetal fibroblasts was similar to those generated by in vitro fertilization. Taken together, our data suggest that the persistent expression of the exogenous factors and the acquisition of chromosomal abnormalities are incompatible with normal development of NT embryos produced with iPSCs. |
| first_indexed | 2025-11-14T19:03:22Z |
| format | Article |
| id | nottingham-28725 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:03:22Z |
| publishDate | 2015 |
| publisher | Mary Ann Liebert |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-287252024-08-15T15:32:51Z https://eprints.nottingham.ac.uk/28725/ Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer German, Sergio D. Campbell, Keith H.S. Thornton, Elisabeth McLachlan, Gerry Sweetman, Dylan Alberio, Ramiro Induced pluripotent stem cells (iPSCs) share similar characteristics of indefinite in vitro growth with embryonic stem cells (ESCs) and may therefore serve as a useful tool for the targeted genetic modification of farm animals via nuclear transfer (NT). Derivation of stable ESC lines from farm animals has not been possible, therefore, it is important to determine whether iPSCs can be used as substitutes for ESCs in generating genetically modified cloned farm animals. We generated ovine iPSCs by conventional retroviral transduction using the four Yamanaka factors. These cells were basic fibroblast growth factor (bFGF)- and activin A-dependent, showed persistent expression of the transgenes, acquired chromosomal abnormalities, and failed to activate endogenous NANOG. Nonetheless, iPSCs could differentiate into the three somatic germ layers in vitro. Because cloning of farm animals is best achieved with diploid cells (G1/G0), we synchronized the iPSCs in G1 prior to NT. Despite the cell cycle synchronization, preimplantation development of iPSC-NT embryos was lower than with somatic cells (2% vs. 10% blastocysts, p<0.01). Furthermore, analysis of the blastocysts produced demonstrated persistent expression of the transgenes, aberrant expression of endogenous SOX2, and a failure to activate NANOG consistently. In contrast, gene expression in blastocysts produced with the parental fetal fibroblasts was similar to those generated by in vitro fertilization. Taken together, our data suggest that the persistent expression of the exogenous factors and the acquisition of chromosomal abnormalities are incompatible with normal development of NT embryos produced with iPSCs. Mary Ann Liebert 2015-02 Article PeerReviewed German, Sergio D., Campbell, Keith H.S., Thornton, Elisabeth, McLachlan, Gerry, Sweetman, Dylan and Alberio, Ramiro (2015) Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer. Cellular Reprogramming, 17 (1). pp. 19-27. ISSN 2152-4971 http://www.liebertpub.com/overview/cellular-reprogramming/9/ doi:10.1089/cell.2014.0071 doi:10.1089/cell.2014.0071 |
| spellingShingle | German, Sergio D. Campbell, Keith H.S. Thornton, Elisabeth McLachlan, Gerry Sweetman, Dylan Alberio, Ramiro Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title | Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title_full | Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title_fullStr | Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title_full_unstemmed | Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title_short | Ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| title_sort | ovine induced pluripotent stem cells are resistant to reprogramming after nuclear transfer |
| url | https://eprints.nottingham.ac.uk/28725/ https://eprints.nottingham.ac.uk/28725/ https://eprints.nottingham.ac.uk/28725/ |