{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T17:13:34Z","timestamp":1790010814817,"version":"4.0.1"},"reference-count":27,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2014,8,23]],"date-time":"2014-08-23T00:00:00Z","timestamp":1408752000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Ministry of Science"},{"name":"ICT & Future Planning","award":["2012M3A9C6049728"],"award-info":[{"award-number":["2012M3A9C6049728"]}]},{"name":"ICT & Future Planning","award":["2012M3A9C6049823"],"award-info":[{"award-number":["2012M3A9C6049823"]}]},{"name":"ICT & Future Planning","award":["2013R1A1A3007591"],"award-info":[{"award-number":["2013R1A1A3007591"]}]},{"DOI":"10.13039\/501100004085","name":"Ministry of Education, Science and Technology","doi-asserted-by":"crossref","award":["2013R1A1A2062781"],"award-info":[{"award-number":["2013R1A1A2062781"]}],"id":[{"id":"10.13039\/501100004085","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100002630","name":"Ewha Womans University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100002630","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Anatomy"],"published-print":{"date-parts":[[2014,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Although mesenchymal stem cells (<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>) isolated from bone marrow and adipose tissues are known to be subjected to <jats:italic>in vitro<\/jats:italic> culture\u2010related alterations in their stem cell properties, such data have not been reported in human tonsil\u2010derived <jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content> (<jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>). Here, we investigated the culture\u2010related changes of phenotypes, the senescence, and the differentiation potential of <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>. <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content> were serially passaged by a standard protocol, and their characteristics were assessed, including <jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>\u2010specific surface antigen profiles, the senescence, and the differentiation potentials into adipocytes, chondrocytes and osteocytes. Up to at least passage 15, we found no alterations in either <jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>\u2010specific surface marker, <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>14, <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>34, <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>45, <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>73 and <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>90, or the m<jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> expression of embryonic stem cell gene markers, <jats:italic><jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>anog<\/jats:italic>,<jats:italic> <jats:styled-content style=\"fixed-case\">O<\/jats:styled-content>ct4\u2010<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content><\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>ox\u20102<\/jats:italic>. However, the expression of <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>146, recently identified another <jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content> marker, dramatically decreased with increasing passages from ~\u00a023% at passage 3 to ~\u00a01% at passage 15. The average doubling time increased significantly from ~\u00a038\u00a0h at passage 10 to ~\u00a046\u00a0h at passage 15. From passage 10, the cell size increased slightly and <jats:styled-content style=\"fixed-case\">SA<\/jats:styled-content>\u2010\u03b2\u2010gal staining was evident. Both <jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>lizarin <jats:styled-content style=\"fixed-case\">R<\/jats:styled-content>ed <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content> staining and osteocalcin expression showed that the osteogenic differentiation potential increased up to passage 10 and decreased thereafter. However, the adipogenic and chondrogenic differentiation potential decreased passage\u2010dependently from the start, as evidenced by staining of <jats:styled-content style=\"fixed-case\">O<\/jats:styled-content>il <jats:styled-content style=\"fixed-case\">R<\/jats:styled-content>ed <jats:styled-content style=\"fixed-case\">O<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>lcian Blue, respectively. Consistent with a passage\u2010dependent osteogenic differentiation, the expression of <jats:styled-content style=\"fixed-case\">CCN<\/jats:styled-content>1, an angiogenic protein known to be related to both senescence and osteogenesis, also increased up to passage 10. Furthermore, ectopic expression of small interfering <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> against <jats:styled-content style=\"fixed-case\">CCN<\/jats:styled-content>1 at passage 10 significantly reversed <jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>lizarin <jats:styled-content style=\"fixed-case\">R<\/jats:styled-content>ed <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content> staining and osteocalcin expression. Altogether, our study demonstrates the characterization of long\u2010term <jats:italic>in vitro<\/jats:italic> cultured <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content> and that <jats:styled-content style=\"fixed-case\">CCN<\/jats:styled-content>1 may be involved in mediating a passage\u2010dependent increase in osteogenic potential of <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MSC<\/jats:styled-content>.<\/jats:p>","DOI":"10.1111\/joa.12229","type":"journal-article","created":{"date-parts":[[2014,8,23]],"date-time":"2014-08-23T08:09:59Z","timestamp":1408781399000},"page":"510-518","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":56,"title":["Characterization of long\u2010term <i>in vitro<\/i> culture\u2010related alterations of human tonsil\u2010derived mesenchymal stem cells: role for <scp>CCN<\/scp>1 in replicative senescence\u2010associated increase in osteogenic differentiation"],"prefix":"10.1111","volume":"225","author":[{"given":"Yeonsil","family":"Yu","sequence":"first","affiliation":[{"name":"Department of Molecular Medicine School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoon Shin","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Han Su","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology\u2010Head &amp; Neck Surgery School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ha Yeong","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoon Mi","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sung\u2010Chul","family":"Jung","sequence":"additional","affiliation":[{"name":"Department of Biochemistry School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyung\u2010Ha","family":"Ryu","sequence":"additional","affiliation":[{"name":"Department of Pediatrics School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inho","family":"Jo","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine School of Medicine, and Global Top 5 Research Program Ewha Womans University Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2014,8,23]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-06-4690"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0142-9612(99)00192-1"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/jcmm.12168"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2007.11.021"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1089\/scd.2010.0040"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4827(65)90211-9"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/ar2459"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1089\/clo.2004.6.369"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tea.2008.0465"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1089\/scd.2008.0335"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0043523"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2121-7-14"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.1174"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s007740050081"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.24179"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1590\/S0103-64402005000300010"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.3109\/14653249.2012.706708"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.26.8.2955-2964.2006"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bone.2003.07.005"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1089\/scd.2011.0722"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.087122"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2004.02.038"},{"key":"e_1_2_8_24_1","first-page":"52","article-title":"The basic properties of mesenchymal stromal cells from the donor bone marrow: superficial markers","volume":"82","author":"Svinareva DA","year":"2010","journal-title":"Ter Arkh"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1089\/107632702753725076"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.4252\/wjsc.v4.i6.53"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0005846"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0002213"}],"container-title":["Journal of Anatomy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fjoa.12229","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fjoa.12229","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/joa.12229","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,2]],"date-time":"2023-10-02T07:08:37Z","timestamp":1696230517000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/joa.12229"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,8,23]]},"references-count":27,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2014,11]]}},"alternative-id":["10.1111\/joa.12229"],"URL":"https:\/\/doi.org\/10.1111\/joa.12229","archive":["Portico"],"relation":{},"ISSN":["0021-8782","1469-7580"],"issn-type":[{"value":"0021-8782","type":"print"},{"value":"1469-7580","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,8,23]]},"assertion":[{"value":"2014-07-23","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-08-23","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}