naa a22 4500 510738982 CHVBK 20180411083021.0 cr unu---uuuuu 180411e20130201xx s 000 0 eng 10.1007/s12015-012-9386-3 doi (NATIONALLICENCE)springer-10.1007/s12015-012-9386-3 Mesenchymal Stromal Cell Phenotype is not Influenced by Confluence during Culture Expansion [Elektronische Daten] [Mandana Haack-Sørensen, Susanne Hansen, Louise Hansen, Michael Gaster, Poul Hyttel, Annette Ekblond, Jens Kastrup] Background: Accumulating preclinical and clinical evidence indicates that human mesenchymal stromal cells (MSCs) are good candidates for cell therapy. For clinical applications of MSCs extensive in vitro expansion is required to obtain an adequate number of cells. It is evident that the pursuit for cell quantity must not affect quality, but it is also a fact that in vitro culture conditions affect MSC phenotype. One possible variable is the degree of cell confluence during expansion. Methods: We investigate the influence of cell density on homogeneity and differentiation during culture expansion of un-stimulated MSCs isolated from the bone marrow in DMEM and fetal bovine serum (FBS). MSC morphology, phenotype and differentiation were investigated weekly during 5weeks culture expansion using electron microscopy, flow cytometry, immunocytochemistry, qualitative RT-PCR and quantitative Q-PCR. Results: The morphological observation and the phenotypic analyses showed that MSCs after 3weeks cultivation constituted a phenotypically homogenous MSC cell population, which at low levels expressed genes for different cell lineages, confirming their multilineage plasticity, without actual differentiation. This phenotype persisted independent of increasing cell densities. Discussion: These data demonstrate that MSC characteristics and plasticity can be maintained during culture expansion from bone marrow mononuclear cells to MSCs and that a homogeneous phenotype of undifferentiated MSCs which persists independent of cell density can be used for clinical therapies. Springer Science+Business Media, LLC, 2012 Bone marrow nationallicence Mesenchymal stromal cell nationallicence Culture expansion nationallicence Spontaneous differentiation nationallicence Clinical use nationallicence Stem cell nationallicence Haack-Sørensen Mandana Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut Hansen Susanne Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut Hansen Louise Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut Gaster Michael Department of Pathology and Department of Endocrinology (KMEB), Odense University Hospital, Odense C, Denmark aut Hyttel Poul Department of Basic Animal and Veterinary Science, Faculty of Life Science, University of Copenhagen, Frederiksberg C, Denmark aut Ekblond Annette Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut Kastrup Jens Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut Stem Cell Reviews and Reports Humana Press Inc 9/1(2013-02-01), 44-58 1550-8943 9:1<44 2013 9 12015 https://doi.org/10.1007/s12015-012-9386-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12015-012-9386-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Haack-Sørensen Mandana Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Hansen Susanne Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Hansen Louise Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Gaster Michael Department of Pathology and Department of Endocrinology (KMEB), Odense University Hospital, Odense C, Denmark aut NATIONALLICENCE 700 1- Hyttel Poul Department of Basic Animal and Veterinary Science, Faculty of Life Science, University of Copenhagen, Frederiksberg C, Denmark aut NATIONALLICENCE 700 1- Ekblond Annette Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Kastrup Jens Cardiology Stem Cell Laboratory, The Heart Centre, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark aut NATIONALLICENCE 773 0- Stem Cell Reviews and Reports Humana Press Inc 9/1(2013-02-01), 44-58 1550-8943 9:1<44 2013 9 12015 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer