Circulating CD34+ cell populations characterized by a low rate (up to five) or high rate (more than five) of cell divisions were isolated from 8 d cultures in the presence of stem cell factor (SCF), interleukin-3 (IL-3), granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), Flt3 ligand and Peg-rHu megakaryocyte growth and development factor (Peg-rHuMGDF) using the fluorescent dye 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometric cell sorting. Phenotypic characterization of cells which had experienced up to five divisions (CFDA-SEbright) showed a similar surface antigen expression to starting, freshly isolated CD34+ cells. Conversely, cells which experienced more than five divisions (CFDA-SEdim) showed a differentiating behaviour, down-regulating CD34 antigen and acquiring differentiation markers. CFDA-SEbright cells were significantly enriched in CD105 (endoglin) positive precursors as compared to both freshly isolated CD34+ and CFDA-SEdim cells. Functional analysis indicated that CFDA-SEbright had a 3-fold and 10-fold greater cumulative cloning efficiency as compared to freshly isolated CD34+ cells and CFDA-SEdim cells, respectively. CFDA-SEbright cells retained the vast majority of LTC-IC and showed a LTC-IC frequency 2.8-fold higher than that found in freshly isolated CD34+ cells. RT-PCR and Western blot analyses showed significantly higher bcl-2 RNA and protein levels in CFDA-SEbright cells as compared to freshly isolated CD34+ and CFDA-SEdim cells. This study indicates that cytokine low-responding circulating CD34+ cells (CFDA-SEbright cells) represent a functionally, phenotypically and molecularly distinct multipotent progenitor population with biological properties associated with primitive precursors.

Functional, phenotypic and molecular characterization of cytokine low-responding circulating CD34+ haemopoietic progenitors / Pierelli, Luca; G., Scambia; A., Fattorossi; G., Bonanno; A., Battaglia; C., Rumi; M., Marone; S., Mozzetti; S., Rutella; G., Menichella; V., Romeo; S., Mancuso; G., Leone. - In: BRITISH JOURNAL OF HAEMATOLOGY. - ISSN 0007-1048. - 102:5(1998), pp. 1139-1150.

Functional, phenotypic and molecular characterization of cytokine low-responding circulating CD34+ haemopoietic progenitors.

PIERELLI, LUCA;
1998

Abstract

Circulating CD34+ cell populations characterized by a low rate (up to five) or high rate (more than five) of cell divisions were isolated from 8 d cultures in the presence of stem cell factor (SCF), interleukin-3 (IL-3), granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), Flt3 ligand and Peg-rHu megakaryocyte growth and development factor (Peg-rHuMGDF) using the fluorescent dye 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometric cell sorting. Phenotypic characterization of cells which had experienced up to five divisions (CFDA-SEbright) showed a similar surface antigen expression to starting, freshly isolated CD34+ cells. Conversely, cells which experienced more than five divisions (CFDA-SEdim) showed a differentiating behaviour, down-regulating CD34 antigen and acquiring differentiation markers. CFDA-SEbright cells were significantly enriched in CD105 (endoglin) positive precursors as compared to both freshly isolated CD34+ and CFDA-SEdim cells. Functional analysis indicated that CFDA-SEbright had a 3-fold and 10-fold greater cumulative cloning efficiency as compared to freshly isolated CD34+ cells and CFDA-SEdim cells, respectively. CFDA-SEbright cells retained the vast majority of LTC-IC and showed a LTC-IC frequency 2.8-fold higher than that found in freshly isolated CD34+ cells. RT-PCR and Western blot analyses showed significantly higher bcl-2 RNA and protein levels in CFDA-SEbright cells as compared to freshly isolated CD34+ and CFDA-SEdim cells. This study indicates that cytokine low-responding circulating CD34+ cells (CFDA-SEbright cells) represent a functionally, phenotypically and molecularly distinct multipotent progenitor population with biological properties associated with primitive precursors.
1998
analysis; antigens; cd; cd34; cell cycle; cell division; cell surface; classification/drug effects/pathology; cultured; cytokines; drug effects; female; fluoresceins; fluorescent dyes; hematopoietic stem cells; humans; immunophenotyping; metabolism; methods; ovarian neoplasms; pathology; pharmacology; proto-oncogene proteins; receptors; succinimides; tumor cells; vascular cell adhesion molecule-1
01 Pubblicazione su rivista::01a Articolo in rivista
Functional, phenotypic and molecular characterization of cytokine low-responding circulating CD34+ haemopoietic progenitors / Pierelli, Luca; G., Scambia; A., Fattorossi; G., Bonanno; A., Battaglia; C., Rumi; M., Marone; S., Mozzetti; S., Rutella; G., Menichella; V., Romeo; S., Mancuso; G., Leone. - In: BRITISH JOURNAL OF HAEMATOLOGY. - ISSN 0007-1048. - 102:5(1998), pp. 1139-1150.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/484166
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