Human umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium
Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are in the foreground as a preferable application for treating diseases. However, the safety of hUC-MSCs after long-term culturing in vitro in serum-free medium remains unclear. hUC-MSCs were separated by adherent tissue culture. hUC-MSC...
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| Veröffentlicht in: | PloS one 2014-06, Vol.9 (6), p.e98565 |
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| description | Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are in the foreground as a preferable application for treating diseases. However, the safety of hUC-MSCs after long-term culturing in vitro in serum-free medium remains unclear.
hUC-MSCs were separated by adherent tissue culture. hUC-MSCs were cultured in serum-free MesenCult-XF medium and FBS-bases DMEM complete medium. At the 1st, 3rd, 5th, 8th, 10th, and 15th passage, the differentiation of MSCs into osteogenic, chondrogenic, and adipogenic cells was detected, and MTT, surface antigens were measured. Tumorigenicity was analyzed at the 15th passage. Conventional karyotyping was performed at passage 0, 8, and 15. The telomerase activity of hUC-MSCs at passage 1-15 was analyzed.
Flow cytometry analysis showed that very high expression was detected for CD105, CD73, and CD90 and very low expression for CD45, CD34, CD14, CD79a, and HLA-DR. MSCs could differentiate into osteocytes, chondrocytes, and adipocytes in vitro. There was no obvious chromosome elimination, displacement, or chromosomal imbalance as determined from the guidelines of the International System for Human Cytogenetic Nomenclature. Telomerase activity was down-regulated significantly when the culture time was prolonged. Further, no tumors formed in rats injected with hUC-MSCs (P15) cultured in serum-free and in serum-containing conditions.
Our data showed that hUC-MSCs met the International Society for Cellular Therapy standards for conditions of long-term in vitro culturing at P15. Since hUC-MSCs can be safely expanded in vitro and are not susceptible to malignant transformation in serum-free medium, these cells are suitable for cell therapy. |
| doi_str_mv | 10.1371/journal.pone.0098565 |
| format | Article |
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hUC-MSCs were separated by adherent tissue culture. hUC-MSCs were cultured in serum-free MesenCult-XF medium and FBS-bases DMEM complete medium. At the 1st, 3rd, 5th, 8th, 10th, and 15th passage, the differentiation of MSCs into osteogenic, chondrogenic, and adipogenic cells was detected, and MTT, surface antigens were measured. Tumorigenicity was analyzed at the 15th passage. Conventional karyotyping was performed at passage 0, 8, and 15. The telomerase activity of hUC-MSCs at passage 1-15 was analyzed.
Flow cytometry analysis showed that very high expression was detected for CD105, CD73, and CD90 and very low expression for CD45, CD34, CD14, CD79a, and HLA-DR. MSCs could differentiate into osteocytes, chondrocytes, and adipocytes in vitro. There was no obvious chromosome elimination, displacement, or chromosomal imbalance as determined from the guidelines of the International System for Human Cytogenetic Nomenclature. Telomerase activity was down-regulated significantly when the culture time was prolonged. Further, no tumors formed in rats injected with hUC-MSCs (P15) cultured in serum-free and in serum-containing conditions.
Our data showed that hUC-MSCs met the International Society for Cellular Therapy standards for conditions of long-term in vitro culturing at P15. Since hUC-MSCs can be safely expanded in vitro and are not susceptible to malignant transformation in serum-free medium, these cells are suitable for cell therapy.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0098565</identifier><identifier>PMID: 24887492</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adipocytes ; Analysis ; Antigens ; Biocompatibility ; Biology and Life Sciences ; Biomedical materials ; Bone marrow ; Cancer ; Cardiology ; CD105 antigen ; CD14 antigen ; CD34 antigen ; CD45 antigen ; CD73 antigen ; CD90 antigen ; Cell culture ; Cell Transformation, Neoplastic ; Cells, Cultured ; Chondrocytes ; Chromosomes ; Culture Media, Serum-Free ; Cytometry ; Data processing ; Flow Cytometry ; Genetic transformation ; Good Manufacturing Practice ; Histocompatibility antigen HLA ; Humans ; Immunophenotyping ; International standardization ; Karyotyping ; Medical research ; Medical treatment ; Medicine and Health Sciences ; Mesenchymal stem cells ; Mesenchymal Stem Cells - cytology ; Mesenchymal Stem Cells - immunology ; Mesenchyme ; Osteocytes ; Rats ; Serum-free medium ; Skin & tissue grafts ; Stem cells ; Studies ; Surface antigens ; Telomerase ; Therapy ; Tissue culture ; Transformation ; Tumorigenicity ; Tumors ; Umbilical cord ; Umbilical Cord - cytology</subject><ispartof>PloS one, 2014-06, Vol.9 (6), p.e98565</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Chen 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>2014 Chen et al 2014 Chen et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-719607d8648adddc6acf258950c228e9dca20213f9f626f10ffcb7d484be983e3</citedby><cites>FETCH-LOGICAL-c692t-719607d8648adddc6acf258950c228e9dca20213f9f626f10ffcb7d484be983e3</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/PMC4041760/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4041760/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24887492$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Ivanovic, Zoran</contributor><creatorcontrib>Chen, Gecai</creatorcontrib><creatorcontrib>Yue, Aihuan</creatorcontrib><creatorcontrib>Ruan, Zhongbao</creatorcontrib><creatorcontrib>Yin, Yigang</creatorcontrib><creatorcontrib>Wang, RuZhu</creatorcontrib><creatorcontrib>Ren, Yin</creatorcontrib><creatorcontrib>Zhu, Li</creatorcontrib><title>Human umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are in the foreground as a preferable application for treating diseases. However, the safety of hUC-MSCs after long-term culturing in vitro in serum-free medium remains unclear.
hUC-MSCs were separated by adherent tissue culture. hUC-MSCs were cultured in serum-free MesenCult-XF medium and FBS-bases DMEM complete medium. At the 1st, 3rd, 5th, 8th, 10th, and 15th passage, the differentiation of MSCs into osteogenic, chondrogenic, and adipogenic cells was detected, and MTT, surface antigens were measured. Tumorigenicity was analyzed at the 15th passage. Conventional karyotyping was performed at passage 0, 8, and 15. The telomerase activity of hUC-MSCs at passage 1-15 was analyzed.
Flow cytometry analysis showed that very high expression was detected for CD105, CD73, and CD90 and very low expression for CD45, CD34, CD14, CD79a, and HLA-DR. MSCs could differentiate into osteocytes, chondrocytes, and adipocytes in vitro. There was no obvious chromosome elimination, displacement, or chromosomal imbalance as determined from the guidelines of the International System for Human Cytogenetic Nomenclature. Telomerase activity was down-regulated significantly when the culture time was prolonged. Further, no tumors formed in rats injected with hUC-MSCs (P15) cultured in serum-free and in serum-containing conditions.
Our data showed that hUC-MSCs met the International Society for Cellular Therapy standards for conditions of long-term in vitro culturing at P15. Since hUC-MSCs can be safely expanded in vitro and are not susceptible to malignant transformation in serum-free medium, these cells are suitable for cell therapy.</description><subject>Adipocytes</subject><subject>Analysis</subject><subject>Antigens</subject><subject>Biocompatibility</subject><subject>Biology and Life Sciences</subject><subject>Biomedical materials</subject><subject>Bone marrow</subject><subject>Cancer</subject><subject>Cardiology</subject><subject>CD105 antigen</subject><subject>CD14 antigen</subject><subject>CD34 antigen</subject><subject>CD45 antigen</subject><subject>CD73 antigen</subject><subject>CD90 antigen</subject><subject>Cell culture</subject><subject>Cell Transformation, Neoplastic</subject><subject>Cells, Cultured</subject><subject>Chondrocytes</subject><subject>Chromosomes</subject><subject>Culture Media, Serum-Free</subject><subject>Cytometry</subject><subject>Data processing</subject><subject>Flow Cytometry</subject><subject>Genetic transformation</subject><subject>Good Manufacturing Practice</subject><subject>Histocompatibility antigen HLA</subject><subject>Humans</subject><subject>Immunophenotyping</subject><subject>International standardization</subject><subject>Karyotyping</subject><subject>Medical research</subject><subject>Medical treatment</subject><subject>Medicine and Health Sciences</subject><subject>Mesenchymal stem cells</subject><subject>Mesenchymal Stem Cells - cytology</subject><subject>Mesenchymal Stem Cells - immunology</subject><subject>Mesenchyme</subject><subject>Osteocytes</subject><subject>Rats</subject><subject>Serum-free medium</subject><subject>Skin & tissue grafts</subject><subject>Stem cells</subject><subject>Studies</subject><subject>Surface antigens</subject><subject>Telomerase</subject><subject>Therapy</subject><subject>Tissue culture</subject><subject>Transformation</subject><subject>Tumorigenicity</subject><subject>Tumors</subject><subject>Umbilical cord</subject><subject>Umbilical 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umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium</title><author>Chen, Gecai ; Yue, Aihuan ; Ruan, Zhongbao ; Yin, Yigang ; Wang, RuZhu ; Ren, Yin ; Zhu, Li</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-719607d8648adddc6acf258950c228e9dca20213f9f626f10ffcb7d484be983e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Adipocytes</topic><topic>Analysis</topic><topic>Antigens</topic><topic>Biocompatibility</topic><topic>Biology and Life Sciences</topic><topic>Biomedical materials</topic><topic>Bone marrow</topic><topic>Cancer</topic><topic>Cardiology</topic><topic>CD105 antigen</topic><topic>CD14 antigen</topic><topic>CD34 antigen</topic><topic>CD45 antigen</topic><topic>CD73 antigen</topic><topic>CD90 antigen</topic><topic>Cell culture</topic><topic>Cell Transformation, 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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>Chen, Gecai</au><au>Yue, Aihuan</au><au>Ruan, Zhongbao</au><au>Yin, Yigang</au><au>Wang, RuZhu</au><au>Ren, Yin</au><au>Zhu, Li</au><au>Ivanovic, Zoran</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Human umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-06-02</date><risdate>2014</risdate><volume>9</volume><issue>6</issue><spage>e98565</spage><pages>e98565-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are in the foreground as a preferable application for treating diseases. However, the safety of hUC-MSCs after long-term culturing in vitro in serum-free medium remains unclear.
hUC-MSCs were separated by adherent tissue culture. hUC-MSCs were cultured in serum-free MesenCult-XF medium and FBS-bases DMEM complete medium. At the 1st, 3rd, 5th, 8th, 10th, and 15th passage, the differentiation of MSCs into osteogenic, chondrogenic, and adipogenic cells was detected, and MTT, surface antigens were measured. Tumorigenicity was analyzed at the 15th passage. Conventional karyotyping was performed at passage 0, 8, and 15. The telomerase activity of hUC-MSCs at passage 1-15 was analyzed.
Flow cytometry analysis showed that very high expression was detected for CD105, CD73, and CD90 and very low expression for CD45, CD34, CD14, CD79a, and HLA-DR. MSCs could differentiate into osteocytes, chondrocytes, and adipocytes in vitro. There was no obvious chromosome elimination, displacement, or chromosomal imbalance as determined from the guidelines of the International System for Human Cytogenetic Nomenclature. Telomerase activity was down-regulated significantly when the culture time was prolonged. Further, no tumors formed in rats injected with hUC-MSCs (P15) cultured in serum-free and in serum-containing conditions.
Our data showed that hUC-MSCs met the International Society for Cellular Therapy standards for conditions of long-term in vitro culturing at P15. Since hUC-MSCs can be safely expanded in vitro and are not susceptible to malignant transformation in serum-free medium, these cells are suitable for cell therapy.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24887492</pmid><doi>10.1371/journal.pone.0098565</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2014-06, Vol.9 (6), p.e98565 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1531473244 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adipocytes Analysis Antigens Biocompatibility Biology and Life Sciences Biomedical materials Bone marrow Cancer Cardiology CD105 antigen CD14 antigen CD34 antigen CD45 antigen CD73 antigen CD90 antigen Cell culture Cell Transformation, Neoplastic Cells, Cultured Chondrocytes Chromosomes Culture Media, Serum-Free Cytometry Data processing Flow Cytometry Genetic transformation Good Manufacturing Practice Histocompatibility antigen HLA Humans Immunophenotyping International standardization Karyotyping Medical research Medical treatment Medicine and Health Sciences Mesenchymal stem cells Mesenchymal Stem Cells - cytology Mesenchymal Stem Cells - immunology Mesenchyme Osteocytes Rats Serum-free medium Skin & tissue grafts Stem cells Studies Surface antigens Telomerase Therapy Tissue culture Transformation Tumorigenicity Tumors Umbilical cord Umbilical Cord - cytology |
| title | Human umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-02T08%3A46%3A24IST&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=Human%20umbilical%20cord-derived%20mesenchymal%20stem%20cells%20do%20not%20undergo%20malignant%20transformation%20during%20long-term%20culturing%20in%20serum-free%20medium&rft.jtitle=PloS%20one&rft.au=Chen,%20Gecai&rft.date=2014-06-02&rft.volume=9&rft.issue=6&rft.spage=e98565&rft.pages=e98565-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0098565&rft_dat=%3Cgale_plos_%3EA416777098%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=1531473244&rft_id=info:pmid/24887492&rft_galeid=A416777098&rft_doaj_id=oai_doaj_org_article_6c80df8371b24b2fbaf80b226a05fa25&rfr_iscdi=true |