title: Pro-B cells propagated in stromal cell-free cultures reconstitute functional B-cell compartments in immunodeficient mice.
abstract: Up to now long-term in vitro growth of pro-B cells was thought to require stromal cells. However, here we show that fetal liver (FL) and bone marrow (BM) derived pro-B cells can be propagated long-term in stromal cell-free cultures supplemented with IL-7, stem cell factor and FLT3 ligand. Within a week, most cells expressed surface CD19, CD79A, λ5, and VpreB antigens and had rearranged immunoglobulin D-J heavy chain genes. Both FL and BM pro-B cells reconstituted the B-cell compartments of immuno-incompetent Rag2-deficient mice, with FL pro-B cells generating follicular, marginal zone (MZB) and B1a B cells, and BM pro-B cells giving rise mainly to MZB cells. Reconstituted Rag2-deficient mice generated significant levels of IgM and IgG antibodies to a type II T-independent antigen; mice reconstituted with FL pro-B cells generated surprisingly high IgG1titers. Finally, we show for the first time that mice reconstituted with mixtures of pro-B and pro-T cells propagated in stromal cell-free in vitro cultures mounted a T-cell-dependent antibody response. This novel stromal cell-free culture system facilitates our understanding of B-cell development and might be applied clinically.
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Lilly
authors.initials: L
authors.lastname: von Muenchow
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Tsapogas
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Llucia
authors.initials: L
authors.lastname: Albertí-Servera
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Capoferri
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.|Molecular Immune Regulation, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Marianne
authors.initials: M
authors.lastname: Doelz
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Hannie
authors.initials: H
authors.lastname: Rolink
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Nabil
authors.initials: N
authors.lastname: Bosco
authors.affiliation: Discipline of Physiology, National University of Ireland, Galway.
authors.forename: Rhodri
authors.initials: R
authors.lastname: Ceredig
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Antonius G
authors.initials: AG
authors.lastname: Rolink
mesh_terms: D000818:Animals; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology*; D001854:Bone Marrow Cells / Q000276:immunology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics; D015851:Interleukin-7 / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D054448:Precursor Cells, B-Lymphoid / Q000276:immunology*; D054504:Precursor Cells, T-Lymphoid / Q000276:immunology; D019089:Stem Cell Factor / Q000378:metabolism; D017154:Stromal Cells / Q000276:immunology; D018183:Transplantation Chimera
chemical_list: D004268:DNA-Binding Proteins; D015851:Interleukin-7; D008565:Membrane Proteins; C499308:Rag2 protein, mouse; D019089:Stem Cell Factor; C084881:flt3 ligand protein
keywords: B cells; Bone marrow; Cell development; Fetal liver; Stroma; Transplantation