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Derivation of Induced Trophoblast Cell Lines in Cattle by Doxycycline-Inducible piggyBac Vectors
Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental...
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Published in: | PloS one 2016-12, Vol.11 (12), p.e0167550-e0167550 |
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creator | Kawaguchi, Takamasa Cho, Dooseon Hayashi, Masafumi Tsukiyama, Tomoyuki Kimura, Koji Matsuyama, Shuichi Minami, Naojiro Yamada, Masayasu Imai, Hiroshi |
description | Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3⁄4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast-related (CDX2, ELF5, ERRβ, and IFN-τ) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy-related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle. |
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The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3⁄4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast-related (CDX2, ELF5, ERRβ, and IFN-τ) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy-related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0167550</identifier><identifier>PMID: 27907214</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amnion ; Amnion - cytology ; Amnion - drug effects ; Amnion - metabolism ; Amniotic cells ; Animals ; Antibiotics ; Beef cattle ; Biology ; Biology and Life Sciences ; Biomarkers - metabolism ; Biotechnology ; c-Myc protein ; Cattle ; CDX2 protein ; Cell culture ; Cell Line ; Cell lineage ; Cell Lineage - drug effects ; Cell lines ; Colonies ; Comparative analysis ; Cytology ; Differentiation ; Dosage and administration ; Doxycycline ; Doxycycline - pharmacology ; Ectoderm - cytology ; Ectoderm - drug effects ; Ectoderm - metabolism ; Embryos ; Female ; Fibroblasts ; Founder Effect ; Gene Expression ; Genes ; Genetic aspects ; Genetic Vectors - chemistry ; Genetic Vectors - metabolism ; Grasslands ; Growth factors ; Implantation ; Induced Pluripotent Stem Cells - cytology ; Induced Pluripotent Stem Cells - drug effects ; Induced Pluripotent Stem Cells - metabolism ; Interferon ; KLF4 protein ; Kruppel-Like Transcription Factors - genetics ; Kruppel-Like Transcription Factors - metabolism ; Laboratories ; Life sciences ; Livestock ; Mammals ; Morphology ; Myc protein ; Oct-4 protein ; Octamer Transcription Factor-3 - genetics ; Octamer Transcription Factor-3 - metabolism ; Placenta ; Pluripotency ; Pregnancy ; Proto-Oncogene Proteins c-myc - genetics ; Proto-Oncogene Proteins c-myc - metabolism ; Research and Analysis Methods ; SOXB1 Transcription Factors - genetics ; SOXB1 Transcription Factors - metabolism ; Spheroids ; Stem cells ; Transcription factors ; Trophectoderm ; Trophoblasts - cytology ; Trophoblasts - drug effects ; Trophoblasts - metabolism</subject><ispartof>PloS one, 2016-12, Vol.11 (12), p.e0167550-e0167550</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 Kawaguchi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2016 Kawaguchi et al 2016 Kawaguchi et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c835t-48e9e17272b1cee0c9b20a246a719f58456ffbf7600046455b73affa4fb8051e3</citedby><cites>FETCH-LOGICAL-c835t-48e9e17272b1cee0c9b20a246a719f58456ffbf7600046455b73affa4fb8051e3</cites><orcidid>0000-0002-0067-5804</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/1845246630/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/1845246630?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27907214$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Knott, Jason Glenn</contributor><creatorcontrib>Kawaguchi, Takamasa</creatorcontrib><creatorcontrib>Cho, Dooseon</creatorcontrib><creatorcontrib>Hayashi, Masafumi</creatorcontrib><creatorcontrib>Tsukiyama, Tomoyuki</creatorcontrib><creatorcontrib>Kimura, Koji</creatorcontrib><creatorcontrib>Matsuyama, Shuichi</creatorcontrib><creatorcontrib>Minami, Naojiro</creatorcontrib><creatorcontrib>Yamada, Masayasu</creatorcontrib><creatorcontrib>Imai, Hiroshi</creatorcontrib><title>Derivation of Induced Trophoblast Cell Lines in Cattle by Doxycycline-Inducible piggyBac Vectors</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3⁄4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast-related (CDX2, ELF5, ERRβ, and IFN-τ) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy-related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.</description><subject>Amnion</subject><subject>Amnion - cytology</subject><subject>Amnion - drug effects</subject><subject>Amnion - metabolism</subject><subject>Amniotic cells</subject><subject>Animals</subject><subject>Antibiotics</subject><subject>Beef cattle</subject><subject>Biology</subject><subject>Biology and Life Sciences</subject><subject>Biomarkers - metabolism</subject><subject>Biotechnology</subject><subject>c-Myc protein</subject><subject>Cattle</subject><subject>CDX2 protein</subject><subject>Cell culture</subject><subject>Cell Line</subject><subject>Cell lineage</subject><subject>Cell Lineage - drug effects</subject><subject>Cell lines</subject><subject>Colonies</subject><subject>Comparative analysis</subject><subject>Cytology</subject><subject>Differentiation</subject><subject>Dosage and administration</subject><subject>Doxycycline</subject><subject>Doxycycline - pharmacology</subject><subject>Ectoderm - cytology</subject><subject>Ectoderm - drug effects</subject><subject>Ectoderm - metabolism</subject><subject>Embryos</subject><subject>Female</subject><subject>Fibroblasts</subject><subject>Founder Effect</subject><subject>Gene Expression</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic Vectors - chemistry</subject><subject>Genetic Vectors - metabolism</subject><subject>Grasslands</subject><subject>Growth factors</subject><subject>Implantation</subject><subject>Induced Pluripotent Stem Cells - cytology</subject><subject>Induced Pluripotent Stem Cells - drug effects</subject><subject>Induced Pluripotent Stem Cells - metabolism</subject><subject>Interferon</subject><subject>KLF4 protein</subject><subject>Kruppel-Like Transcription Factors - genetics</subject><subject>Kruppel-Like Transcription Factors - metabolism</subject><subject>Laboratories</subject><subject>Life sciences</subject><subject>Livestock</subject><subject>Mammals</subject><subject>Morphology</subject><subject>Myc protein</subject><subject>Oct-4 protein</subject><subject>Octamer Transcription Factor-3 - genetics</subject><subject>Octamer Transcription Factor-3 - metabolism</subject><subject>Placenta</subject><subject>Pluripotency</subject><subject>Pregnancy</subject><subject>Proto-Oncogene Proteins c-myc - genetics</subject><subject>Proto-Oncogene Proteins c-myc - metabolism</subject><subject>Research and Analysis Methods</subject><subject>SOXB1 Transcription Factors - genetics</subject><subject>SOXB1 Transcription Factors - metabolism</subject><subject>Spheroids</subject><subject>Stem cells</subject><subject>Transcription factors</subject><subject>Trophectoderm</subject><subject>Trophoblasts - cytology</subject><subject>Trophoblasts - drug effects</subject><subject>Trophoblasts - metabolism</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNqNk11v0zAUhiMEYmPwDxBEQkJw0eJvJzdIo-OjUqVJMHZrHMdOPblxsZNp_fc4azY1aBeTL2z5PO_r42OfLHsNwRxiDj9d-T600s23vtVzABmnFDzJjmGJ0YwhgJ8erI-yFzFeAUBxwdjz7AjxEnAEyXH250wHey0769vcm3zZ1r3SdX4R_HbtKydjly-0c_nKtjrmts0XsuuczqtdfuZvdmqnXIrMbnW2SoGtbZrdF6nyS606H-LL7JmRLupX43yS_f729WLxY7Y6_75cnK5mqsC0m5FClxpyxFEFldZAlRUCEhEmOSwNLQhlxlSGMwAAYYTSimNpjCSmKgCFGp9kb_e-W-ejGIsTBUzK5MIwSMRyT9ReXoltsBsZdsJLK243fGiEDJ1VTgtJFQBMlWWhKSmNlIinwxXmuGYQU5S8Po-n9dVG10q3XZBuYjqNtHYtGn8tKMQIA5IMPowGwf_tdezExkaVKi1b7fshb1ZgzAEvHoESWiBK2JDWu__QhwsxUo1Md7Wt8SlFNZiKU8IRhqCkAzV_gEqj1hur0qczNu1PBB8ngsR0-qZrZB-jWP76-Xj2_HLKvj9g11q6bh2964c_G6cg2YMq-BiDNvfvAYEYeuauGmLoGTH2TJK9OXzLe9Fdk-B_HCEP4A</recordid><startdate>20161201</startdate><enddate>20161201</enddate><creator>Kawaguchi, Takamasa</creator><creator>Cho, Dooseon</creator><creator>Hayashi, Masafumi</creator><creator>Tsukiyama, Tomoyuki</creator><creator>Kimura, Koji</creator><creator>Matsuyama, Shuichi</creator><creator>Minami, Naojiro</creator><creator>Yamada, Masayasu</creator><creator>Imai, Hiroshi</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-0067-5804</orcidid></search><sort><creationdate>20161201</creationdate><title>Derivation of Induced Trophoblast Cell Lines in Cattle by Doxycycline-Inducible piggyBac Vectors</title><author>Kawaguchi, Takamasa ; Cho, Dooseon ; Hayashi, Masafumi ; Tsukiyama, Tomoyuki ; Kimura, Koji ; Matsuyama, Shuichi ; Minami, Naojiro ; Yamada, Masayasu ; Imai, Hiroshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c835t-48e9e17272b1cee0c9b20a246a719f58456ffbf7600046455b73affa4fb8051e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Amnion</topic><topic>Amnion - cytology</topic><topic>Amnion - drug effects</topic><topic>Amnion - metabolism</topic><topic>Amniotic cells</topic><topic>Animals</topic><topic>Antibiotics</topic><topic>Beef cattle</topic><topic>Biology</topic><topic>Biology and Life Sciences</topic><topic>Biomarkers - metabolism</topic><topic>Biotechnology</topic><topic>c-Myc protein</topic><topic>Cattle</topic><topic>CDX2 protein</topic><topic>Cell culture</topic><topic>Cell Line</topic><topic>Cell lineage</topic><topic>Cell Lineage - drug effects</topic><topic>Cell lines</topic><topic>Colonies</topic><topic>Comparative analysis</topic><topic>Cytology</topic><topic>Differentiation</topic><topic>Dosage and administration</topic><topic>Doxycycline</topic><topic>Doxycycline - pharmacology</topic><topic>Ectoderm - cytology</topic><topic>Ectoderm - drug effects</topic><topic>Ectoderm - metabolism</topic><topic>Embryos</topic><topic>Female</topic><topic>Fibroblasts</topic><topic>Founder Effect</topic><topic>Gene Expression</topic><topic>Genes</topic><topic>Genetic aspects</topic><topic>Genetic Vectors - chemistry</topic><topic>Genetic Vectors - metabolism</topic><topic>Grasslands</topic><topic>Growth factors</topic><topic>Implantation</topic><topic>Induced Pluripotent Stem Cells - cytology</topic><topic>Induced Pluripotent Stem Cells - drug effects</topic><topic>Induced Pluripotent Stem Cells - metabolism</topic><topic>Interferon</topic><topic>KLF4 protein</topic><topic>Kruppel-Like Transcription Factors - genetics</topic><topic>Kruppel-Like Transcription Factors - metabolism</topic><topic>Laboratories</topic><topic>Life sciences</topic><topic>Livestock</topic><topic>Mammals</topic><topic>Morphology</topic><topic>Myc protein</topic><topic>Oct-4 protein</topic><topic>Octamer Transcription Factor-3 - genetics</topic><topic>Octamer Transcription Factor-3 - metabolism</topic><topic>Placenta</topic><topic>Pluripotency</topic><topic>Pregnancy</topic><topic>Proto-Oncogene Proteins c-myc - genetics</topic><topic>Proto-Oncogene Proteins c-myc - metabolism</topic><topic>Research and Analysis Methods</topic><topic>SOXB1 Transcription Factors - genetics</topic><topic>SOXB1 Transcription Factors - metabolism</topic><topic>Spheroids</topic><topic>Stem cells</topic><topic>Transcription factors</topic><topic>Trophectoderm</topic><topic>Trophoblasts - cytology</topic><topic>Trophoblasts - drug effects</topic><topic>Trophoblasts - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kawaguchi, Takamasa</creatorcontrib><creatorcontrib>Cho, Dooseon</creatorcontrib><creatorcontrib>Hayashi, Masafumi</creatorcontrib><creatorcontrib>Tsukiyama, Tomoyuki</creatorcontrib><creatorcontrib>Kimura, Koji</creatorcontrib><creatorcontrib>Matsuyama, Shuichi</creatorcontrib><creatorcontrib>Minami, Naojiro</creatorcontrib><creatorcontrib>Yamada, Masayasu</creatorcontrib><creatorcontrib>Imai, Hiroshi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale_Opposing Viewpoints In Context</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>ProQuest_Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection (Proquest) (PQ_SDU_P3)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kawaguchi, Takamasa</au><au>Cho, Dooseon</au><au>Hayashi, Masafumi</au><au>Tsukiyama, Tomoyuki</au><au>Kimura, Koji</au><au>Matsuyama, Shuichi</au><au>Minami, Naojiro</au><au>Yamada, Masayasu</au><au>Imai, Hiroshi</au><au>Knott, Jason Glenn</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Derivation of Induced Trophoblast Cell Lines in Cattle by Doxycycline-Inducible piggyBac Vectors</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2016-12-01</date><risdate>2016</risdate><volume>11</volume><issue>12</issue><spage>e0167550</spage><epage>e0167550</epage><pages>e0167550-e0167550</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3⁄4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast-related (CDX2, ELF5, ERRβ, and IFN-τ) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy-related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27907214</pmid><doi>10.1371/journal.pone.0167550</doi><tpages>e0167550</tpages><orcidid>https://orcid.org/0000-0002-0067-5804</orcidid><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1932-6203 |
ispartof | PloS one, 2016-12, Vol.11 (12), p.e0167550-e0167550 |
issn | 1932-6203 1932-6203 |
language | eng |
recordid | cdi_plos_journals_1845246630 |
source | PubMed (Medline); ProQuest - Publicly Available Content Database |
subjects | Amnion Amnion - cytology Amnion - drug effects Amnion - metabolism Amniotic cells Animals Antibiotics Beef cattle Biology Biology and Life Sciences Biomarkers - metabolism Biotechnology c-Myc protein Cattle CDX2 protein Cell culture Cell Line Cell lineage Cell Lineage - drug effects Cell lines Colonies Comparative analysis Cytology Differentiation Dosage and administration Doxycycline Doxycycline - pharmacology Ectoderm - cytology Ectoderm - drug effects Ectoderm - metabolism Embryos Female Fibroblasts Founder Effect Gene Expression Genes Genetic aspects Genetic Vectors - chemistry Genetic Vectors - metabolism Grasslands Growth factors Implantation Induced Pluripotent Stem Cells - cytology Induced Pluripotent Stem Cells - drug effects Induced Pluripotent Stem Cells - metabolism Interferon KLF4 protein Kruppel-Like Transcription Factors - genetics Kruppel-Like Transcription Factors - metabolism Laboratories Life sciences Livestock Mammals Morphology Myc protein Oct-4 protein Octamer Transcription Factor-3 - genetics Octamer Transcription Factor-3 - metabolism Placenta Pluripotency Pregnancy Proto-Oncogene Proteins c-myc - genetics Proto-Oncogene Proteins c-myc - metabolism Research and Analysis Methods SOXB1 Transcription Factors - genetics SOXB1 Transcription Factors - metabolism Spheroids Stem cells Transcription factors Trophectoderm Trophoblasts - cytology Trophoblasts - drug effects Trophoblasts - metabolism |
title | Derivation of Induced Trophoblast Cell Lines in Cattle by Doxycycline-Inducible piggyBac Vectors |
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