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dc.contributor.authorRossi, Barbara
dc.contributor.authorZanolin, Elisabetta
dc.contributor.authorVincenzi, Carlo
dc.contributor.authorDiani, Franco
dc.contributor.authorPizzolo, Giovanni
dc.contributor.authorDe Wynter, Erika A
dc.contributor.authorNadali, Gianpaolo
dc.date.accessioned2009-08-25T11:15:11Z
dc.date.available2009-08-25T11:15:11Z
dc.date.issued2004-08
dc.identifier.citationEffect of addition of FLT-3 ligand and megakaryocyte growth and development factor on hemopoietic cells in serum-free conditions. 2004, 13 (4):362-71 Stem Cells Dev.en
dc.identifier.issn1547-3287
dc.identifier.pmid15345130
dc.identifier.doi10.1089/1547328041797453
dc.identifier.urihttp://hdl.handle.net/10541/78446
dc.description.abstractThe aim of this study was to clarify the mechanisms that regulate hematopoietic cell expansion in vitro by identifying defined culture conditions. We report the results of experiments with CD34(+) cells from cord blood (CB, n = 13), bone marrow (BM, n = 4), and mobilized peripheral blood stem cells (PBSC, n = 5) using two combinations of cytokines: (A) granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), stem cell factor (SCF), erythropoietin (EPO), insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (FGF-b) and (B) combination A plus FLT3 ligand (FL) and megakaryocyte growth and development factor (PEG rhMGDF). Cultures of immunoselected CD34(+) cells were performed in serum-free liquid medium without serum substitutes. The area under the curve (AUC) obtained by plotting the logarithm of the total number of viable cells, CD34(+) cells, and CFC per well, toward the week of culture was used as an index of cell expansion. With CB, a significant difference was obtained between the two combinations of cytokines with regard to the total number of viable cells, GM-CFC, and CD34(+) cells. The difference between the two combinations of cytokines obtained with BM was significant with respect to the total number of viable cells and CD34(+) cells but not for the erythroid and myeloid progenitors. When CD34(+) cells from peripheral blood stem cells (PBSC) were cultured in presence of the two combinations of cytokines, the difference in terms of AUC was not statistically significant. Our data indicate additional effects in terms of proliferation and expansion of hematopoietic cells in serum-free conditions when FL and polyethylene glycol (PEG) rhMGDF are included in culture and suggest a differential activity of these cytokines on cells from different hematopoietic sources.
dc.language.isoenen
dc.subjectHaematopoiesisen
dc.subjectHaematopoietic Stem Cellsen
dc.subjectHaematopoietic Stem Cell Mobilisationen
dc.subject.meshAntigens, CD
dc.subject.meshAntigens, CD34
dc.subject.meshBone Marrow Cells
dc.subject.meshCell Culture Techniques
dc.subject.meshCell Separation
dc.subject.meshCulture Media, Serum-Free
dc.subject.meshFetal Blood
dc.subject.meshFlow Cytometry
dc.subject.meshHematopoiesis
dc.subject.meshHematopoietic Stem Cell Mobilization
dc.subject.meshHematopoietic Stem Cells
dc.subject.meshHumans
dc.subject.meshInfant, Newborn
dc.subject.meshMembrane Proteins
dc.subject.meshThrombopoietin
dc.titleEffect of addition of FLT-3 ligand and megakaryocyte growth and development factor on hemopoietic cells in serum-free conditions.en
dc.typeArticleen
dc.contributor.departmentDepartment of Clinical and Experimental Medicine, Section of Hematology, University of Verona, Italy.en
dc.identifier.journalStem Cells and Developmenten
html.description.abstractThe aim of this study was to clarify the mechanisms that regulate hematopoietic cell expansion in vitro by identifying defined culture conditions. We report the results of experiments with CD34(+) cells from cord blood (CB, n = 13), bone marrow (BM, n = 4), and mobilized peripheral blood stem cells (PBSC, n = 5) using two combinations of cytokines: (A) granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), stem cell factor (SCF), erythropoietin (EPO), insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (FGF-b) and (B) combination A plus FLT3 ligand (FL) and megakaryocyte growth and development factor (PEG rhMGDF). Cultures of immunoselected CD34(+) cells were performed in serum-free liquid medium without serum substitutes. The area under the curve (AUC) obtained by plotting the logarithm of the total number of viable cells, CD34(+) cells, and CFC per well, toward the week of culture was used as an index of cell expansion. With CB, a significant difference was obtained between the two combinations of cytokines with regard to the total number of viable cells, GM-CFC, and CD34(+) cells. The difference between the two combinations of cytokines obtained with BM was significant with respect to the total number of viable cells and CD34(+) cells but not for the erythroid and myeloid progenitors. When CD34(+) cells from peripheral blood stem cells (PBSC) were cultured in presence of the two combinations of cytokines, the difference in terms of AUC was not statistically significant. Our data indicate additional effects in terms of proliferation and expansion of hematopoietic cells in serum-free conditions when FL and polyethylene glycol (PEG) rhMGDF are included in culture and suggest a differential activity of these cytokines on cells from different hematopoietic sources.


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