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Accordingly, we noticed that a significant number of these cells, which are smaller than erythrocytes, are lost during gradient centrifugation over Ficoll\u2010Paque as well as during routine volume depletion of UCB units before freezing. To preserve these cells in final UCB preparations, we propose a relatively short and economical three\u2010step isolation protocol that allows recovery of approximately 60% of the initial number of Lin<jats:sup>\u2212<\/jats:sup>\/CD45<jats:sup>\u2212<\/jats:sup>\/CD133<jats:sup>+<\/jats:sup> UCB\u2010VSELs present in freshly harvested UCB units. In this novel approach (i) UCB is lysed in a hypotonic ammonium chloride solution to deplete erythrocytes; (ii) CD133<jats:sup>+<\/jats:sup> including VSELs cells are enriched by employing immunomagnetic beads; and subsequently (iii) Lin<jats:sup>\u2212<\/jats:sup>\/CD45<jats:sup>\u2212<\/jats:sup>\/CD133<jats:sup>+<\/jats:sup> cells are sorted by fluorescence\u2010activated cell sorting. The whole isolation procedure takes approximately 2\u20133\u2003h per UCB unit and isolated cells are highly enriched for an Oct\u20104<jats:sup>+<\/jats:sup> and SSEA\u20104<jats:sup>+<\/jats:sup> population of small Lin<jats:sup>\u2212<\/jats:sup>\/CD45<jats:sup>\u2212<\/jats:sup>\/CD133<jats:sup>+<\/jats:sup> cells.<\/jats:p>","DOI":"10.1111\/j.1600-0609.2009.01352.x","type":"journal-article","created":{"date-parts":[[2009,9,15]],"date-time":"2009-09-15T02:17:40Z","timestamp":1252981060000},"page":"34-46","source":"Crossref","is-referenced-by-count":53,"title":["Optimization of isolation and further characterization of umbilical cord blood\u2010derived very small embryonic\/ epiblast\u2010like stem cells (VSELs)"],"prefix":"10.1111","volume":"84","author":[{"given":"Ewa 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