Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line
Recently, numerous studies have identified that immature cell populations including stem cells and progenitor cells can be found among "side-population" (SP) cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trop...
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| Veröffentlicht in: | PloS one 2011-07, Vol.6 (7), p.e21990 |
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| creator | Takao, Tomoka Asanoma, Kazuo Kato, Kiyoko Fukushima, Kotaro Tsunematsu, Ryosuke Hirakawa, Toshio Matsumura, Sueo Seki, Hiroyuki Takeda, Satoru Wake, Norio |
| description | Recently, numerous studies have identified that immature cell populations including stem cells and progenitor cells can be found among "side-population" (SP) cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population. |
| doi_str_mv | 10.1371/journal.pone.0021990 |
| format | Article |
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Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0021990</identifier><identifier>PMID: 21760941</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Animals ; Antigens ; Biology ; Biomarkers ; Biomarkers - metabolism ; Cancer therapies ; Cell Line ; Cell Proliferation - drug effects ; Cell Separation - methods ; Cells (biology) ; Colonies ; Conditioning ; Culture Media, Conditioned - pharmacology ; Cytokines ; Differentiation ; DNA microarrays ; Down-Regulation - drug effects ; Embryo fibroblasts ; Embryo, Mammalian - cytology ; Female ; Fetuses ; Fibroblast growth factor 2 ; Fibroblasts - cytology ; Fibroblasts - drug effects ; Fibroblasts - metabolism ; Gene expression ; Genetic aspects ; Gynecology ; Heparin ; Humans ; Intelligent design ; Interleukin 1 ; Interleukin 7 receptors ; Mice ; Obstetrics ; Placenta ; Population ; Pregnancy ; Pregnancy Trimester, First - drug effects ; Receptors, Interleukin-1 - metabolism ; Receptors, Interleukin-7 - metabolism ; Set-top boxes (Television) ; Side-Population Cells - cytology ; Side-Population Cells - drug effects ; Side-Population Cells - metabolism ; Stem cell transplantation ; Stem cells ; Tissues ; Trophoblasts - cytology ; Trophoblasts - drug effects ; Trophoblasts - metabolism ; Up-Regulation - drug effects</subject><ispartof>PloS one, 2011-07, Vol.6 (7), p.e21990</ispartof><rights>COPYRIGHT 2011 Public Library of Science</rights><rights>2011 Takao et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://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. 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Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. 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Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>21760941</pmid><doi>10.1371/journal.pone.0021990</doi><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_plos_journals_1305139013 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Public Library of Science (PLoS); PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Animals Antigens Biology Biomarkers Biomarkers - metabolism Cancer therapies Cell Line Cell Proliferation - drug effects Cell Separation - methods Cells (biology) Colonies Conditioning Culture Media, Conditioned - pharmacology Cytokines Differentiation DNA microarrays Down-Regulation - drug effects Embryo fibroblasts Embryo, Mammalian - cytology Female Fetuses Fibroblast growth factor 2 Fibroblasts - cytology Fibroblasts - drug effects Fibroblasts - metabolism Gene expression Genetic aspects Gynecology Heparin Humans Intelligent design Interleukin 1 Interleukin 7 receptors Mice Obstetrics Placenta Population Pregnancy Pregnancy Trimester, First - drug effects Receptors, Interleukin-1 - metabolism Receptors, Interleukin-7 - metabolism Set-top boxes (Television) Side-Population Cells - cytology Side-Population Cells - drug effects Side-Population Cells - metabolism Stem cell transplantation Stem cells Tissues Trophoblasts - cytology Trophoblasts - drug effects Trophoblasts - metabolism Up-Regulation - drug effects |
| title | Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-19T02%3A09%3A57IST&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=Isolation%20and%20characterization%20of%20human%20trophoblast%20side-population%20(SP)%20cells%20in%20primary%20villous%20cytotrophoblasts%20and%20HTR-8/SVneo%20cell%20line&rft.jtitle=PloS%20one&rft.au=Takao,%20Tomoka&rft.date=2011-07-07&rft.volume=6&rft.issue=7&rft.spage=e21990&rft.pages=e21990-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0021990&rft_dat=%3Cgale_plos_%3EA476886309%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=1305139013&rft_id=info:pmid/21760941&rft_galeid=A476886309&rft_doaj_id=oai_doaj_org_article_4db254af05de43dbab60454446de29c4&rfr_iscdi=true |