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To this end, we sorted out of the freshly established ASCs four subpopulations (SPs) according to a specific pattern of co-expression of six surface markers, the CD34, CD73, CD90, CD105, CD146, and CD271, using polychromatic flow cytometry.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Method<\/jats:title>\n                <jats:p>Using flow cytometry-associated cell sorting and analysis, gating parameters were set to select for a CD73<jats:sup>+<\/jats:sup>CD90<jats:sup>+<\/jats:sup>CD105<jats:sup>+<\/jats:sup> phenotype plus one of the four following combinations, CD34<jats:sup>\u2212<\/jats:sup>CD146<jats:sup>\u2212<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> (SP1), CD34<jats:sup>\u2212<\/jats:sup>CD146<jats:sup>+<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> (SP2), CD34<jats:sup>+<\/jats:sup>CD146<jats:sup>+<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> (SP3), and CD34<jats:sup>\u2212<\/jats:sup>CD146<jats:sup>+<\/jats:sup>CD271<jats:sup>+<\/jats:sup> (SP4). The SPs were expanded 700- to 1000-fold, and their surface repertoire, trilineage differentiation, and clonogenic potential, and the capacity to support wound healing were assayed.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Upon culturing, the co-expression of major epitopes, the CD73, CD90, and CD105 was maintained, while regarding the minor markers, all SPs reverted to resemble the pre-sorted population with CD34<jats:sup>\u2212<\/jats:sup>CD146<jats:sup>\u2212<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> and CD34<jats:sup>\u2212<\/jats:sup>CD146<jats:sup>+<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> representing the most prevalent combinations, followed by less frequent CD34<jats:sup>+<\/jats:sup>CD146<jats:sup>\u2212<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> and CD34<jats:sup>+<\/jats:sup>CD146<jats:sup>+<\/jats:sup>CD271<jats:sup>\u2212<\/jats:sup> variants. There was no difference in the efficiency of adipo-, osteo-, or chondrogenesis by cytochemistry and real-time RT-PCR or the CFU capacity between the individual SPs, however, the SP2<jats:sup>CD73+90+105+34-146+271-<\/jats:sup> outperformed others in terms of wound healing.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Our study shows that ASCs upon culturing inherently maintain a stable distribution of immunophenotype variants, which may potentially disguise specific functional properties of particular downstream lines. Furthermore, the outlined approach suggests a paradigm whereby discrete subpopulations could be identified to provide for a therapeutically most relevant cell product.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s13287-016-0435-8","type":"journal-article","created":{"date-parts":[[2016,12,1]],"date-time":"2016-12-01T12:32:07Z","timestamp":1480595527000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Discrete adipose-derived stem cell subpopulations may display differential functionality after in vitro expansion despite convergence to a common phenotype distribution"],"prefix":"10.1186","volume":"7","author":[{"given":"Frederik M\u00f8lgaard","family":"Nielsen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simone Elkj\u00e6r","family":"Riis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jens Isak","family":"Andersen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rapha\u00eblle","family":"Lesage","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Trine","family":"Fink","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristian Pablo","family":"Pennisi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vladimir","family":"Zachar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2016,12,1]]},"reference":[{"key":"435_CR1","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1089\/107632701300062859","volume":"7","author":"PA Zuk","year":"2001","unstructured":"Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. 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