CD34 human hematopoietic progenitor cell line, MUTZ-3, differentiates into functional osteoclasts
| Autor: | Donatella Barisani, Anna Berardi, E. Ciraci, Antonina Parafioriti, Giuseppe Formisano, Gianfranco Bottazzo, Giuseppe Arancia |
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| Přispěvatelé: | Ciraci, E, Barisani, D, Parafioriti, A, Formisano, G, Arancia, G, Bottazzo, G, Berardi, A |
| Rok vydání: | 2007 |
| Předmět: |
Macrophage colony-stimulating factor
Cancer Research Time Factors Cell Population Lipopolysaccharide Receptors CD34 Osteoclasts Antigens CD34 Receptor Macrophage Colony-Stimulating Factor Biology Monocytes Cell Line Osteoclast Genetics medicine Humans education Molecular Biology education.field_of_study Receptor Activator of Nuclear Factor-kappa B Monocyte osteoclast differentiation stem cells BIO/13 - BIOLOGIA APPLICATA Cell Biology Hematology Hematopoietic Stem Cells Antigens Differentiation Cell biology medicine.anatomical_structure Cell culture Immunology Microscopy Electron Scanning Cytokines Stem cell |
| Zdroj: | Experimental Hematology. 35:967-977 |
| ISSN: | 0301-472X |
| DOI: | 10.1016/j.exphem.2007.03.003 |
| Popis: | Objective. CD14(+) monocyte cell lines can differentiate into an osteoclast (OC)-like lineage. However, the identification of human cell lines with stem cell characteristics, capable of differentiating into OCs, would provide a tool for the study of the molecular mechanisms regulating their commitment, differentiation, and function. Since the human acute myeloid leukemia cell line MUTZ-3 contains both CD34(+) stem cell and CD14(+) cell populations, we investigated the capacity of the stem/progenitor CD34(+) population to differentiate into functional OCs. Materials and Methods. Sorted MUTZ-3-CD34(+) and MUTZ-3-CD14(+) cells were cultured in presence of M-CSF, RANK-L, and TNF-alpha to generate OCs. Differentiation was evaluated by TRAP staining and RT-PCR, which assessed the expression of c-fms, RANK, MMP-9, CATK, TRAP, and CTR in -CD34(+)OC and -CD14(+)OC cells. Resorption pit formation was also evaluated. CD34, CD14, M-CSF-R, RANK, and CTR expression was assessed by FACS analysis. Results. MUTZ-3-CD34(+) differentiated into OCs, displaying the full range of differentiation markers; MMP-9, CATK, TRAP, and RANK mRNA were detected from day 3 of culture, whereas CTR from day 12. Stimulated MUTZ-3-CD34(+) generated functional osteoclasts that formed extensive resorption lacunae on both mineralized surface and bone slices. Surprisingly, in both sorted populations we identified a population M-CSF-R+/RANK(+) that at the same time co-expressed CD14 and CD34. Conclusions. These findings demonstrate that MUTZ-3 cells constitute an invaluable model to study the expression pattern in different developmental stages of commitment and differentiation. Importantly, the data indicate that the CD14(+)CD34(+)M-CSF-R(+)RANK(+) population represents an intermediate stage of differentiation from CD34 precursors and monocytes to osteoclast. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc. |
| Databáze: | OpenAIRE |
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