Bovine trophectoderm cells induced from bovine fibroblasts with induced pluripotent stem cell reprogramming factors

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dc.contributor.authorTalbot, Neil C.en
dc.contributor.authorSparks, Wendy O.en
dc.contributor.authorPhillips, Caitlin E.en
dc.contributor.authorEaly, Alan D.en
dc.contributor.authorPowell, Anne M.en
dc.contributor.authorCaperna, Thomas J.en
dc.contributor.authorGarrett, Wesley M.en
dc.contributor.authorDonovan, David M.en
dc.contributor.authorBlomberg, Le Annen
dc.contributor.departmentAnimal and Poultry Sciencesen
dc.date.accessioned2020-03-23T13:26:00Zen
dc.date.available2020-03-23T13:26:00Zen
dc.date.issued2017-06en
dc.description.abstractThirteen independent induced bovine trophectroderm (iBT) cell lines were established by reprogramming bovine fetal liver-derived fibroblasts after viral-vector transduction with either six or eight factors, including POU5F1 (OCT4), KLF4, SOX2, MYC, NANOG, LIN28, SV40 large T antigen, and hTERT. Light-and electron-microscopy analysis showed that the iBT cells had epithelial cell morphology typical of bovine trophectoderm cells. Reverse-transcription-PCR assays indicated that all of the cell lines expressed interferon-tau (IFNT) at passages 1 or 2. At later passages (>= passage 8), however, immunoblot and antiviral activity assays revealed that more than half of the iBT cell lines had stopped expressing IFNT. Messenger RNAs specific to trophectoderm differentiation and function were found in the iBT cell lines, and 2-dimensional gel analysis for cellular proteins showed an expression pattern similar to that of trophectoderm cell lines derived from bovine blastocysts. Integration of some of the human reprogramming factors, including POU5F1, KLF4, SOX2, MYC, NANOG, and LIN28, were detected by PCR, but their transcription was mostly absent in the iBT cell lines. Gene expression assessment of endogenous bovine reprogramming factor orthologs revealed endogenous bLIN28 and bMYC transcripts in all; bSOX2 and bNANOG in none; and bKLF4 and bPOU5F1 in less than half of the iBT cell lines. These results demonstrate that bovine trophectoderm can be induced via reprogramming factor expression from bovine liver-derived fibroblasts, although other fibroblast populations-e.g., derived from fetal thigh tissue-may produce similar results, albeit at lower frequencies.en
dc.description.adminPublic domain – authored by a U.S. government employeeen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1002/mrd.22797en
dc.identifier.eissn1098-2795en
dc.identifier.issn1040-452Xen
dc.identifier.issue6en
dc.identifier.pmid28332752en
dc.identifier.urihttp://hdl.handle.net/10919/97436en
dc.identifier.volume84en
dc.language.isoenen
dc.rightsCC0 1.0 Universalen
dc.rights.urihttp://creativecommons.org/publicdomain/zero/1.0/en
dc.subjectbovineen
dc.subjectcell cultureen
dc.subjectinduceden
dc.subjectiPSCen
dc.subjectreprogrammingen
dc.subjecttrophectodermen
dc.titleBovine trophectoderm cells induced from bovine fibroblasts with induced pluripotent stem cell reprogramming factorsen
dc.title.serialMolecular Reproduction and Developmenten
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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