An Evaluation of the Stemness, Paracrine, and Tumorigenic Characteristics of Highly Expanded, Minimally Passaged Adipose-Derived Stem Cells
The use of adipose-derived stem cells (ADSC) in regenerative medicine is rising due to their plasticity, capacity of differentiation and paracrine and trophic effects. Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases...
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| Veröffentlicht in: | PloS one 2016-09, Vol.11 (9), p.e0162332 |
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| creator | El Atat, Oula Antonios, Diane Hilal, George Hokayem, Nabil Abou-Ghoch, Joelle Hashim, Hussein Serhal, Rim Hebbo, Clara Moussa, Mayssam Alaaeddine, Nada |
| description | The use of adipose-derived stem cells (ADSC) in regenerative medicine is rising due to their plasticity, capacity of differentiation and paracrine and trophic effects. Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases or cosmetic procedures. Thus, there is the need for culturing and expanding cells in-vitro for several weeks remain. Our aim is to investigate if long- term proliferation with minimal passaging will affect the stemness, paracrine secretions and carcinogenesis markers of ADSC. The immunophenotypic properties and aldehyde dehydrogenase (ALDH) activity of the initial stromal vascular fraction (SVF) and serially passaged ADSC were observed by flow cytometry. In parallel, the telomerase activity and the relative expression of oncogenes and tumor suppressor genes were assessed by q-PCR. We also assessed the cytokine secretion profile of passaged ADSC by an ELISA. The expanded ADSC retain their morphological and phenotypical characteristics. These cells maintained in culture for up to 12 weeks until P4, possessed stable telomerase and ALDH activity, without having a TP53 mutation. Furthermore, the relative expression levels of TP53, RB, and MDM2 were not affected while the relative expression of c-Myc decreased significantly. Finally, the levels of the secretions of PGE2, STC1, and TIMP2 were not affected but the levels of IL-6, VEGF, and TIMP 1 significantly decreased at P2. Our results suggest that the expansion of passaged ADSC does not affect the differentiation capacity of stem cells and does not confer a cancerous state or capacity in vitro to the cells. |
| doi_str_mv | 10.1371/journal.pone.0162332 |
| format | Article |
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Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases or cosmetic procedures. Thus, there is the need for culturing and expanding cells in-vitro for several weeks remain. Our aim is to investigate if long- term proliferation with minimal passaging will affect the stemness, paracrine secretions and carcinogenesis markers of ADSC. The immunophenotypic properties and aldehyde dehydrogenase (ALDH) activity of the initial stromal vascular fraction (SVF) and serially passaged ADSC were observed by flow cytometry. In parallel, the telomerase activity and the relative expression of oncogenes and tumor suppressor genes were assessed by q-PCR. We also assessed the cytokine secretion profile of passaged ADSC by an ELISA. The expanded ADSC retain their morphological and phenotypical characteristics. These cells maintained in culture for up to 12 weeks until P4, possessed stable telomerase and ALDH activity, without having a TP53 mutation. Furthermore, the relative expression levels of TP53, RB, and MDM2 were not affected while the relative expression of c-Myc decreased significantly. Finally, the levels of the secretions of PGE2, STC1, and TIMP2 were not affected but the levels of IL-6, VEGF, and TIMP 1 significantly decreased at P2. Our results suggest that the expansion of passaged ADSC does not affect the differentiation capacity of stem cells and does not confer a cancerous state or capacity in vitro to the cells.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0162332</identifier><identifier>PMID: 27632538</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adipocytes ; Adipose tissue ; Adipose Tissue - cytology ; Adipose Tissue - immunology ; Adolescent ; Adult ; Aldehyde dehydrogenase ; Aldehyde Dehydrogenase - metabolism ; Analysis ; Apoptosis ; Biology and Life Sciences ; Bone marrow ; c-Myc protein ; Cancer ; Carcinogenesis ; Carcinogens ; Cell culture ; Cell Differentiation ; Cell Lineage ; Cytometry ; Dehydrogenases ; Dermatology ; Diabetes ; Differentiation ; Enzyme-linked immunosorbent assay ; Fibroblasts ; Flow cytometry ; Gene expression ; Health aspects ; Humans ; Immunophenotyping ; Inflammation ; Interleukin 6 ; Kinases ; Laboratories ; MDM2 protein ; Medicine ; Medicine and Health Sciences ; Mesenchymal stem cells ; Middle Aged ; Mutation ; Myc protein ; p53 Protein ; Paracrine signalling ; Physical characteristics ; Plastic surgery ; Prostaglandin E2 ; Regenerative medicine ; Research and Analysis Methods ; Secretions ; Stem cells ; Stem Cells - cytology ; Stem Cells - immunology ; Telomerase ; Telomerase - metabolism ; Therapeutic applications ; Tissue engineering ; Tissue inhibitor of metalloproteinase 2 ; Tumor proteins ; Tumor suppressor genes ; Tumor Suppressor Protein p53 - metabolism ; Vascular endothelial growth factor ; Young Adult</subject><ispartof>PloS one, 2016-09, Vol.11 (9), p.e0162332</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 El Atat et al. 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cytology</subject><subject>Stem Cells - immunology</subject><subject>Telomerase</subject><subject>Telomerase - metabolism</subject><subject>Therapeutic applications</subject><subject>Tissue engineering</subject><subject>Tissue inhibitor of metalloproteinase 2</subject><subject>Tumor proteins</subject><subject>Tumor suppressor genes</subject><subject>Tumor Suppressor Protein p53 - metabolism</subject><subject>Vascular endothelial growth factor</subject><subject>Young Adult</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk99u0zAUxiMEYmPwBggiTUIgtSW28_cGqSqFVRraxAa31ql9nHpK7GIn1fYMvDQO7aYW7QLlIvHx7_uOz4lPFL0myYSwgny8sb0z0EzW1uAkITlljD6JjknF6DinCXu6930UvfD-JkkyVub58-iIFjmjYXEc_Z6aeL6BpodOWxNbFXcrjK86bA16P4ovwYFw2uAoBiPj6761TtdotIhnq2GvQ6d9p4UftGe6XjV38fx2HWCUo_ibNrqFJsQuwXuoUcZTqdfW4_hzEG7CesgVz7Bp_MvomYLG46vd-yT68WV-PTsbn198Xcym52NR0KwbK1mCKFiZSIYyk2XKoJASEJQQghUpK1m2VESWDGCJQGie5kQJKSumaJIRdhK93fquG-v5ro-ek5JUVVKkWR6IxZaQFm742oUa3B23oPnfgHU1BxeKbpBTJjKRK5WUiqUV0KWEKk-zCjAFAmTI9mmXrV-2KAWazkFzYHq4Y_SK13bDs4SmVTUYvN8ZOPurR9_xVnsRGgYGbT-cmxZlyVhWBvT0H_Tx6nZUDaEAbZQNecVgyqdpkVQpoTQN1OQRKjwSWy3CpVM6xA8EHw4Egenwtquh954vrr7_P3vx85B9t8euEJpu5W3TDxfWH4LpFhTOeu9QPTSZJHyYmftu8GFm-G5mguzN_g96EN0PCfsDqg4TKA</recordid><startdate>20160915</startdate><enddate>20160915</enddate><creator>El Atat, Oula</creator><creator>Antonios, Diane</creator><creator>Hilal, George</creator><creator>Hokayem, Nabil</creator><creator>Abou-Ghoch, Joelle</creator><creator>Hashim, Hussein</creator><creator>Serhal, Rim</creator><creator>Hebbo, Clara</creator><creator>Moussa, Mayssam</creator><creator>Alaaeddine, Nada</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>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20160915</creationdate><title>An Evaluation of the Stemness, Paracrine, and Tumorigenic Characteristics of Highly Expanded, Minimally Passaged Adipose-Derived Stem Cells</title><author>El Atat, Oula ; Antonios, Diane ; Hilal, George ; Hokayem, Nabil ; Abou-Ghoch, Joelle ; Hashim, Hussein ; Serhal, Rim ; Hebbo, Clara ; Moussa, Mayssam ; Alaaeddine, Nada</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c725t-fd8ac7380d3ed5d843a7ddaeafccc3743835bf1d83aabea126461fcdd93f20513</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Adipocytes</topic><topic>Adipose tissue</topic><topic>Adipose Tissue - 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cytology</topic><topic>Stem Cells - immunology</topic><topic>Telomerase</topic><topic>Telomerase - metabolism</topic><topic>Therapeutic applications</topic><topic>Tissue engineering</topic><topic>Tissue inhibitor of metalloproteinase 2</topic><topic>Tumor proteins</topic><topic>Tumor suppressor genes</topic><topic>Tumor Suppressor Protein p53 - metabolism</topic><topic>Vascular endothelial growth factor</topic><topic>Young Adult</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>El Atat, Oula</creatorcontrib><creatorcontrib>Antonios, Diane</creatorcontrib><creatorcontrib>Hilal, George</creatorcontrib><creatorcontrib>Hokayem, Nabil</creatorcontrib><creatorcontrib>Abou-Ghoch, Joelle</creatorcontrib><creatorcontrib>Hashim, Hussein</creatorcontrib><creatorcontrib>Serhal, Rim</creatorcontrib><creatorcontrib>Hebbo, Clara</creatorcontrib><creatorcontrib>Moussa, Mayssam</creatorcontrib><creatorcontrib>Alaaeddine, Nada</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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Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases or cosmetic procedures. Thus, there is the need for culturing and expanding cells in-vitro for several weeks remain. Our aim is to investigate if long- term proliferation with minimal passaging will affect the stemness, paracrine secretions and carcinogenesis markers of ADSC. The immunophenotypic properties and aldehyde dehydrogenase (ALDH) activity of the initial stromal vascular fraction (SVF) and serially passaged ADSC were observed by flow cytometry. In parallel, the telomerase activity and the relative expression of oncogenes and tumor suppressor genes were assessed by q-PCR. We also assessed the cytokine secretion profile of passaged ADSC by an ELISA. The expanded ADSC retain their morphological and phenotypical characteristics. These cells maintained in culture for up to 12 weeks until P4, possessed stable telomerase and ALDH activity, without having a TP53 mutation. Furthermore, the relative expression levels of TP53, RB, and MDM2 were not affected while the relative expression of c-Myc decreased significantly. Finally, the levels of the secretions of PGE2, STC1, and TIMP2 were not affected but the levels of IL-6, VEGF, and TIMP 1 significantly decreased at P2. Our results suggest that the expansion of passaged ADSC does not affect the differentiation capacity of stem cells and does not confer a cancerous state or capacity in vitro to the cells.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27632538</pmid><doi>10.1371/journal.pone.0162332</doi><tpages>e0162332</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2016-09, Vol.11 (9), p.e0162332 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1819907456 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Adipocytes Adipose tissue Adipose Tissue - cytology Adipose Tissue - immunology Adolescent Adult Aldehyde dehydrogenase Aldehyde Dehydrogenase - metabolism Analysis Apoptosis Biology and Life Sciences Bone marrow c-Myc protein Cancer Carcinogenesis Carcinogens Cell culture Cell Differentiation Cell Lineage Cytometry Dehydrogenases Dermatology Diabetes Differentiation Enzyme-linked immunosorbent assay Fibroblasts Flow cytometry Gene expression Health aspects Humans Immunophenotyping Inflammation Interleukin 6 Kinases Laboratories MDM2 protein Medicine Medicine and Health Sciences Mesenchymal stem cells Middle Aged Mutation Myc protein p53 Protein Paracrine signalling Physical characteristics Plastic surgery Prostaglandin E2 Regenerative medicine Research and Analysis Methods Secretions Stem cells Stem Cells - cytology Stem Cells - immunology Telomerase Telomerase - metabolism Therapeutic applications Tissue engineering Tissue inhibitor of metalloproteinase 2 Tumor proteins Tumor suppressor genes Tumor Suppressor Protein p53 - metabolism Vascular endothelial growth factor Young Adult |
| title | An Evaluation of the Stemness, Paracrine, and Tumorigenic Characteristics of Highly Expanded, Minimally Passaged Adipose-Derived Stem Cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-11T04%3A09%3A46IST&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=An%20Evaluation%20of%20the%20Stemness,%20Paracrine,%20and%20Tumorigenic%20Characteristics%20of%20Highly%20Expanded,%20Minimally%20Passaged%20Adipose-Derived%20Stem%20Cells&rft.jtitle=PloS%20one&rft.au=El%20Atat,%20Oula&rft.date=2016-09-15&rft.volume=11&rft.issue=9&rft.spage=e0162332&rft.pages=e0162332-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0162332&rft_dat=%3Cgale_plos_%3EA470941224%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=1819907456&rft_id=info:pmid/27632538&rft_galeid=A470941224&rft_doaj_id=oai_doaj_org_article_23c5c6ff08f349a2bda96459ae4a1a11&rfr_iscdi=true |