Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning
We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and 12.8% vs....
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| Veröffentlicht in: | PloS one 2015-02, Vol.10 (2), p.e0118165-e0118165 |
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| creator | Siriboon, Chawalit Lin, Yu-Hsuan Kere, Michel Chen, Chun-Da Chen, Lih-Ren Chen, Chien-Hong Tu, Ching-Fu Lo, Neng-Wen Ju, Jyh-Cherng |
| description | We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and 12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P < 0.05) in those derived from TSA-treated 3× blastocysts (36.7 and 26.7%) than from the non-treated aggregated group (23.1 and 11.5%). These cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state. |
| doi_str_mv | 10.1371/journal.pone.0118165 |
| format | Article |
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After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and 12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P < 0.05) in those derived from TSA-treated 3× blastocysts (36.7 and 26.7%) than from the non-treated aggregated group (23.1 and 11.5%). These cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0118165</identifier><identifier>PMID: 25680105</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Agricultural biotechnology ; Alkaline phosphatase ; Animal sciences ; Animals ; Apoptosis ; Biomarkers ; Biotechnology ; Blastocyst - cytology ; Blastocysts ; Blastomeres - cytology ; Bovidae ; Cell culture ; Cell Differentiation ; Cell Line ; Cell lines ; Cell proliferation ; Cell survival ; Cells, Cultured ; Cloning ; Cloning, Organism ; Colonies ; Colonization ; Comparative analysis ; Culture Media ; Efficiency ; Embryo cells ; Embryo Culture Techniques ; Embryoid Bodies - cytology ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - metabolism ; Embryos ; Feeder Cells ; Feeders ; Female ; Hogs ; Medical research ; Oct-4 protein ; Oocytes - cytology ; Stem cells ; Suidae ; Survival ; Swine ; Zoology</subject><ispartof>PloS one, 2015-02, Vol.10 (2), p.e0118165-e0118165</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Siriboon 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>2015 Siriboon et al 2015 Siriboon et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-9e8935c1551019eb1416f1685ef3a7bebdb0f4632af85b8440ca03e733f5fb413</citedby><cites>FETCH-LOGICAL-c692t-9e8935c1551019eb1416f1685ef3a7bebdb0f4632af85b8440ca03e733f5fb413</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4334543/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4334543/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2100,2926,23865,27923,27924,53790,53792,79371,79372</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25680105$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Cooney, Austin John</contributor><creatorcontrib>Siriboon, Chawalit</creatorcontrib><creatorcontrib>Lin, Yu-Hsuan</creatorcontrib><creatorcontrib>Kere, Michel</creatorcontrib><creatorcontrib>Chen, Chun-Da</creatorcontrib><creatorcontrib>Chen, Lih-Ren</creatorcontrib><creatorcontrib>Chen, Chien-Hong</creatorcontrib><creatorcontrib>Tu, Ching-Fu</creatorcontrib><creatorcontrib>Lo, Neng-Wen</creatorcontrib><creatorcontrib>Ju, Jyh-Cherng</creatorcontrib><title>Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and 12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P < 0.05) in those derived from TSA-treated 3× blastocysts (36.7 and 26.7%) than from the non-treated aggregated group (23.1 and 11.5%). These cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state.</description><subject>Agricultural biotechnology</subject><subject>Alkaline phosphatase</subject><subject>Animal sciences</subject><subject>Animals</subject><subject>Apoptosis</subject><subject>Biomarkers</subject><subject>Biotechnology</subject><subject>Blastocyst - cytology</subject><subject>Blastocysts</subject><subject>Blastomeres - cytology</subject><subject>Bovidae</subject><subject>Cell culture</subject><subject>Cell Differentiation</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>Cell proliferation</subject><subject>Cell survival</subject><subject>Cells, Cultured</subject><subject>Cloning</subject><subject>Cloning, Organism</subject><subject>Colonies</subject><subject>Colonization</subject><subject>Comparative analysis</subject><subject>Culture Media</subject><subject>Efficiency</subject><subject>Embryo cells</subject><subject>Embryo Culture Techniques</subject><subject>Embryoid Bodies - cytology</subject><subject>Embryonic stem cells</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - metabolism</subject><subject>Embryos</subject><subject>Feeder Cells</subject><subject>Feeders</subject><subject>Female</subject><subject>Hogs</subject><subject>Medical research</subject><subject>Oct-4 protein</subject><subject>Oocytes - cytology</subject><subject>Stem cells</subject><subject>Suidae</subject><subject>Survival</subject><subject>Swine</subject><subject>Zoology</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk12L1DAUhoso7jr6D0QLgujFjPme9kZYFj8GFlb8ug1petLJ0DZjki7Orb_cdKa7zMheSKFNT57zJudNTpY9x2iB6RK_27jB96pdbF0PC4RxgQV_kJ3jkpK5IIg-PBqfZU9C2CDEaSHE4-yMcFEgjPh59ufLEFW0N5Bvnde2hxy6yu9cb3UeInS5hrbN2zQR8hp8AuvceNflqmk8NCqO_2kn85FLY92m7dSTSMi33tWDToFqlzundxHyygbQ0bp-z9q-eZo9MqoN8Gz6zrIfHz98v_w8v7r-tLq8uJprUZI4L6EoKdeYc4xwCRVmWBgsCg6GqmUFVV0hwwQlyhS8KhhDWiEKS0oNNxXDdJa9POhuWxfkZF-QyTYhyqJAJBGrA1E7tZFbbzvld9IpK_cB5xupfLS6BZkUBS4LUQuqGFdYGUpLpYhhNeME66T1flptqDqoNfTRq_ZE9HSmt2vZuBvJKGU8vWbZm0nAu18DhCg7G0aXVQ9uOOwbMY7QWNmrf9D7q5uoRqUCbG9cWlePovKCEYKIIHuXFvdQ6amhszodrrEpfpLw9iQhMRF-x0YNIcjVt6__z17_PGVfH7FrUG1cB9cO49UJpyA7gNq7EDyYO5MxkmOr3Lohx1aRU6uktBfHB3SXdNsb9C9gRA_k</recordid><startdate>20150213</startdate><enddate>20150213</enddate><creator>Siriboon, Chawalit</creator><creator>Lin, Yu-Hsuan</creator><creator>Kere, Michel</creator><creator>Chen, Chun-Da</creator><creator>Chen, Lih-Ren</creator><creator>Chen, Chien-Hong</creator><creator>Tu, Ching-Fu</creator><creator>Lo, Neng-Wen</creator><creator>Ju, Jyh-Cherng</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>AEUYN</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>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150213</creationdate><title>Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning</title><author>Siriboon, Chawalit ; Lin, Yu-Hsuan ; Kere, Michel ; Chen, Chun-Da ; Chen, Lih-Ren ; Chen, Chien-Hong ; Tu, Ching-Fu ; Lo, Neng-Wen ; Ju, Jyh-Cherng</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-9e8935c1551019eb1416f1685ef3a7bebdb0f4632af85b8440ca03e733f5fb413</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Agricultural biotechnology</topic><topic>Alkaline phosphatase</topic><topic>Animal sciences</topic><topic>Animals</topic><topic>Apoptosis</topic><topic>Biomarkers</topic><topic>Biotechnology</topic><topic>Blastocyst - cytology</topic><topic>Blastocysts</topic><topic>Blastomeres - cytology</topic><topic>Bovidae</topic><topic>Cell culture</topic><topic>Cell Differentiation</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>Cell proliferation</topic><topic>Cell survival</topic><topic>Cells, Cultured</topic><topic>Cloning</topic><topic>Cloning, Organism</topic><topic>Colonies</topic><topic>Colonization</topic><topic>Comparative analysis</topic><topic>Culture Media</topic><topic>Efficiency</topic><topic>Embryo cells</topic><topic>Embryo Culture Techniques</topic><topic>Embryoid Bodies - cytology</topic><topic>Embryonic stem cells</topic><topic>Embryonic Stem Cells - cytology</topic><topic>Embryonic Stem Cells - metabolism</topic><topic>Embryos</topic><topic>Feeder Cells</topic><topic>Feeders</topic><topic>Female</topic><topic>Hogs</topic><topic>Medical research</topic><topic>Oct-4 protein</topic><topic>Oocytes - cytology</topic><topic>Stem cells</topic><topic>Suidae</topic><topic>Survival</topic><topic>Swine</topic><topic>Zoology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Siriboon, Chawalit</creatorcontrib><creatorcontrib>Lin, Yu-Hsuan</creatorcontrib><creatorcontrib>Kere, Michel</creatorcontrib><creatorcontrib>Chen, Chun-Da</creatorcontrib><creatorcontrib>Chen, Lih-Ren</creatorcontrib><creatorcontrib>Chen, Chien-Hong</creatorcontrib><creatorcontrib>Tu, Ching-Fu</creatorcontrib><creatorcontrib>Lo, Neng-Wen</creatorcontrib><creatorcontrib>Ju, Jyh-Cherng</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 In Context: Opposing Viewpoints</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>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 Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>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 Korea</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</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>Siriboon, Chawalit</au><au>Lin, Yu-Hsuan</au><au>Kere, Michel</au><au>Chen, Chun-Da</au><au>Chen, Lih-Ren</au><au>Chen, Chien-Hong</au><au>Tu, Ching-Fu</au><au>Lo, Neng-Wen</au><au>Ju, Jyh-Cherng</au><au>Cooney, Austin John</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2015-02-13</date><risdate>2015</risdate><volume>10</volume><issue>2</issue><spage>e0118165</spage><epage>e0118165</epage><pages>e0118165-e0118165</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and 12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P < 0.05) in those derived from TSA-treated 3× blastocysts (36.7 and 26.7%) than from the non-treated aggregated group (23.1 and 11.5%). These cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25680105</pmid><doi>10.1371/journal.pone.0118165</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2015-02, Vol.10 (2), p.e0118165-e0118165 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1656698802 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS); EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Agricultural biotechnology Alkaline phosphatase Animal sciences Animals Apoptosis Biomarkers Biotechnology Blastocyst - cytology Blastocysts Blastomeres - cytology Bovidae Cell culture Cell Differentiation Cell Line Cell lines Cell proliferation Cell survival Cells, Cultured Cloning Cloning, Organism Colonies Colonization Comparative analysis Culture Media Efficiency Embryo cells Embryo Culture Techniques Embryoid Bodies - cytology Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Embryos Feeder Cells Feeders Female Hogs Medical research Oct-4 protein Oocytes - cytology Stem cells Suidae Survival Swine Zoology |
| title | Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-13T00%3A11%3A10IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Putative%20porcine%20embryonic%20stem%20cell%20lines%20derived%20from%20aggregated%20four-celled%20cloned%20embryos%20produced%20by%20oocyte%20bisection%20cloning&rft.jtitle=PloS%20one&rft.au=Siriboon,%20Chawalit&rft.date=2015-02-13&rft.volume=10&rft.issue=2&rft.spage=e0118165&rft.epage=e0118165&rft.pages=e0118165-e0118165&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0118165&rft_dat=%3Cgale_plos_%3EA422026241%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1656698802&rft_id=info:pmid/25680105&rft_galeid=A422026241&rft_doaj_id=oai_doaj_org_article_fb461986d63a45a1af339aa2f4d4521c&rfr_iscdi=true |