Isern, JoanMartín-Antonio, BeatrizGhazanfari, RoshanakMartín, Aan M.López, Juan A.Toro, Raquel delSánchez-Aguilera, AbelArranz, LorenaMartín-Pérez, DanielSuárez-Lledó, MaríaMarín, PedroVan Pel, MelissaFibbe, Willem E.Vázquez, JesúsScheding, StefanUrbano Ispizua, ÁlvaroMéndez-Ferrer, Simón2014-03-102014-03-102013-04-252211-1247https://hdl.handle.net/2445/51084Strategies 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.11 p.application/pdfengcc-by (c) Isern, J. et al., 2013http://creativecommons.org/licenses/by/3.0/esDiferenciació cel·lularHematopoesiMedul·la òssiaCell diferentiationHematopoiesisBone marrowinfo:eu-repo/semantics/article6266932014-03-10info:eu-repo/semantics/openAccess23623496