title: Self-renewing human bone marrow mesenspheres promote hematopoietic stem cell expansion.
abstract: Strategies 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.
authors.affiliation: Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid 28029, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Isern
authors.forename: Beatriz
authors.initials: B
authors.lastname: Martín-Antonio
authors.forename: Roshanak
authors.initials: R
authors.lastname: Ghazanfari
authors.forename: Ana M
authors.initials: AM
authors.lastname: Martín
authors.forename: Juan A
authors.initials: JA
authors.lastname: López
authors.forename: Raquel
authors.initials: R
authors.lastname: del Toro
authors.forename: Abel
authors.initials: A
authors.lastname: Sánchez-Aguilera
authors.forename: Lorena
authors.initials: L
authors.lastname: Arranz
authors.forename: Daniel
authors.initials: D
authors.lastname: Martín-Pérez
authors.forename: María
authors.initials: M
authors.lastname: Suárez-Lledó
authors.forename: Pedro
authors.initials: P
authors.lastname: Marín
authors.forename: Melissa
authors.initials: M
authors.lastname: Van Pel
authors.forename: Willem E
authors.initials: WE
authors.lastname: Fibbe
authors.forename: Jesús
authors.initials: J
authors.lastname: Vázquez
authors.forename: Stefan
authors.initials: S
authors.lastname: Scheding
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Urbano-Ispizúa
authors.forename: Simón
authors.initials: S
authors.lastname: Méndez-Ferrer
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D018920:Coculture Techniques; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D064231:Nestin / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D064231:Nestin