Effect of addition of FLT-3 ligand and megakaryocyte growth and development factor on hemopoietic cells in serum-free conditions.

2.50
Hdl Handle:
http://hdl.handle.net/10541/78446
Title:
Effect of addition of FLT-3 ligand and megakaryocyte growth and development factor on hemopoietic cells in serum-free conditions.
Authors:
Rossi, Barbara; Zanolin, Elisabetta; Vincenzi, Carlo; Diani, Franco; Pizzolo, Giovanni; De Wynter, Erika A; Nadali, Gianpaolo
Abstract:
The 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.
Affiliation:
Department of Clinical and Experimental Medicine, Section of Hematology, University of Verona, Italy.
Citation:
Effect 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.
Journal:
Stem Cells and Development
Issue Date:
Aug-2004
URI:
http://hdl.handle.net/10541/78446
DOI:
10.1089/1547328041797453
PubMed ID:
15345130
Type:
Article
Language:
en
ISSN:
1547-3287
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
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.en
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

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