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C. Carlo-Stella, L. Mangoni, G. Piovani, D. Garau, C. Almici, V. Rizzoli, Biological and chemical selection of ph-negative clones, Stem Cells, Volume 11, Issue S3, 1993, Pages 77–82, https://doi.org/10.1002/stem.5530110919
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Abstract
Chronic myelogenous leukemia (CML) is a clonal disorder of the hematopoietic stem cell characterized by the co-existence of Philadelphia-negative with Ph-positive progenitors. CML progenitor cells have been shown to be defective in adherence to marrow stroma. The present study investigated at the cytogenetic level marrow-derived CML clonogenic cells generated from the stroma-adherent cell fraction. Mononuclear marrow cells from CML patients (n = 20) were incubated with mafosfamide (100 μg/ml) or control medium, seeded onto marrow stromal layers, and allowed to adhere (2 h, 37°C). Following a short-term (3 days) liquid culture, the cells were harvested, incorporated in methylcellulose, and individual colonies were analyzed by single colony karyotyping. On direct cytogenetic analysis, the overall mean (± SD) percentage of Ph-negative metaphases was 9 ± 20%. The mean (± SD) percentages of Ph-negative colonies grown from the stroma-adherent and the stroma-adherent mafosfamide-treated fraction were 41 ± 32% and 62 ± 40% (p ≤ .005), respectively. Single colony transfer experiments revealed that 50 ± 13% stroma-adherent and 70 ± 24% stroma-adherent mafosfamide-treated progenitors gave rise to secondary colonies. In conclusion, the present data demonstrate the possibility to select Ph-negative clones that: 1) have a maintained capability of stroma adherence; 2) are mafosfamide resistant; and 3) have high-replating potential.
