Isern, JoanMartín-Antonio, BeatrizGhazanfari, RoshanakMartin, Ana M.Lopez, Juan AntonioDel Toro, RaquelSanchez-Aguilera, AbelArranz, LorenaMartin-Perez, DanielSuárez-Lledó, MaríaMarín, PedroVan Pel, MelissaFibbe, Willem EVazquez, JesusScheding, StefanUrbano-Ispizúa, ÁlvaroMendez-Ferrer, Simon2018-11-202018-11-202013-05-30Cell Rep. 2013 May 30;3(5):1714-2422111247http://hdl.handle.net/20.500.12105/6643Strategies for expanding hematopoietic stem cells (HSCs) include coculture with cells that recapitulate their natural microenvironment, such as bone marrow stromal stem/progenitor cells (BMSCs). Plastic-adherent BMSCs may be insufficient to preserve primitive HSCs. Here, we describe a method of isolating and culturing human BMSCs as nonadherent mesenchymal spheres. Human mesenspheres were derived from CD45- CD31- CD71- CD146+ CD105+ nestin+ cells but could also be simply grown from fetal and adult BM CD45--enriched cells. Human mesenspheres robustly differentiated into mesenchymal lineages. In culture conditions where they displayed a relatively undifferentiated phenotype, with decreased adherence to plastic and increased self-renewal, they promoted enhanced expansion of cord blood CD34+ cells through secreted soluble factors. Expanded HSCs were serially transplantable in immunodeficient mice and significantly increased long-term human hematopoietic engraftment. These results pave the way for culture techniques that preserve the self-renewal of human BMSCs and their ability to support functional HSCs.engVoRhttp://creativecommons.org/licenses/by-nc-nd/4.0/AnimalsAntigens, CDBone Marrow CellsCell DifferentiationCell LineageCells, CulturedCoculture TechniquesFetal BloodHematopoietic Stem CellsHumansMesenchymal Stromal CellsMiceMice, Inbred NODNestinAttribution-NonCommercial-NoDerivatives 4.0 Internacional23623496351714-2410.1016/j.celrep.2013.03.0412211-1247Cell reportsopen access