Application of growth factor stimulants improves cytogenetic analysis of chronic myeloproliferative disorder patients without alteration to cell lineage or clonality
Autor: | J.D. Howard, Elisabeth P. Nacheva, Beverly Vaughan, M.A. Scott |
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Rok vydání: | 2007 |
Předmět: |
Adult
Male Cancer Research Myeloid Mitotic index medicine.medical_treatment Cellular differentiation Biology Stem cell marker Immunophenotyping Tumor Cells Cultured Genetics medicine Humans Cell Lineage Growth Substances Molecular Biology Aged Aged 80 and over Myeloproliferative Disorders medicine.diagnostic_test Growth factor Middle Aged Molecular biology Clone Cells medicine.anatomical_structure Chronic Disease Cytogenetic Analysis Immunology Female Bone marrow Fluorescence in situ hybridization |
Zdroj: | Cancer Genetics and Cytogenetics. 175:98-106 |
ISSN: | 0165-4608 |
DOI: | 10.1016/j.cancergencyto.2007.02.001 |
Popis: | Conventional cytogenetic methods rely on culturing bone marrow aspirates to obtain suitable and sufficient mitotic figures for G-banded analysis. Samples from patients with chronic myeloproliferative disorders (CMPD) often have increased failure rates due to reduced growth and poor morphology, all of which hamper the conventional karyotyping investigation. The application of growth factor (GF) stimulants to bone marrow aspirates has been shown to yield significant increases in both the quality and quantity of bone marrow metaphases obtained in 53 CPMD patient samples. All cultures were stimulated using the conditioned supernatant from the human bladder carcinoma cell line 5637, which contains IL-3, IL-6, and G-CSF. Results were assessed qualitatively on G-banded preparations and quantitatively by mitotic index (MI = % dividing cells). To assess whether the application of GF stimulants leads to clonal selection, culture samples from 15 patients were analyzed by fluorescence in situ hybridization, which supported the theory that clonal selection remains unaltered in GF-stimulated cultures. In addition to this immunophenotyping of cells, we demonstrated the lineage of cells propagated under these conditions. Cell markers were chosen to characterize B-lymphoid, T-lymphoid, myeloid, and primitive cell types. Results indicated that T cells were maintained in culture and B-lymphoid markers remained negative. In the myeloid subset, there was an overall reduction in the pan-myeloid markers. We believe this represents the loss of terminally differentiated cells (e.g., neutrophils) in culture. Overall, the study clearly demonstrates that the application of GF stimulants does not alter clonality or cell lineage propagated in these samples and is therefore suitable for application in diagnostic cytogenetic laboratories. |
Databáze: | OpenAIRE |
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