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Identification of Philadelphia-negative granulocyte-macrophage colony-forming units generated by stroma-adherent cells from chronic myelogenous leukemia patients.

PIOVANI, Giovanna;
1994-01-01
1994
Altra università italiana
LS7_6 Gene therapy, stem cell therapy, regenerative medicine
BLOOD
2-s2.0-0028044780
WOS:A1994MZ28900026
Inglese
Internazionale
83
1373
1380
7
7509656
Chronic myelogenous leukemia (CML) is a clonal disorder of the hematopoietic stem cell characterized by the coexistence of Philadelphia-negative (Ph-) with Ph+ 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. On direct cytogenetic analysis, the overall mean (+/- SEM) percentage of Ph- metaphases was 3\% +/- 1\%. Mononuclear marrow cells from CML patients (n = 18) were incubated with mafosfamide (100 micrograms/mL) or control medium, seeded onto marrow stromal layers and allowed to adhere (2 hours, 37 degrees C). After a short-term (3-day) liquid culture, the cells were harvested, incorporated in methyl-cellulose, and individual colonies were analyzed by single colony karyotyping. The mean (+/- SEM) percentage of Ph- colonies generated from the stroma-adherent fraction was 35\% +/- 6\%. As compared with marrow colony-forming unit granulocyte-macrophage plated before any manipulation, the mean (+/- SEM) percentage of Ph- clones was significantly increased by stroma adherence (35\% +/- 6\% v 15\% +/- 4\%, P < or = .005) and mafosfamide (100 micrograms/mL) incubation of marrow cells before stroma adherence (58\% +/- 9\% v 35\% +/- 6\%, P < or = .005). An additive effect was observed by combining mafosfamide treatment and stroma adherence. Single-colony transfer experiments showed that 50\% +/- 4\% stroma-adherent and 70\% +/- 4\% stroma-adherent mafosfamide-treated progenitors gave rise to secondary colonies. To further characterize the stroma-adherent fraction, experiments were performed in which CD34+ marrow cells were used. The mean (+/- SEM) output of progenitors generated by 10,000 CD34+, stroma-adherent cells was 888 +/- 188 and 570 +/- 258 for untreated and mafosfamide-treated cells, respectively. Individual colonies were analyzed by single-colony karyotyping and fluorescent in situ hybridization using a biotinylated cosmid DNA probe that hybridize to abl oncogene. The CD34+, stroma-adherent fraction contained 38\% +/- 14\% (untreated) and 56\% +/- 18\% (mafosfamide-treated) (P < or = .025) Ph- progenitors. In conclusion, the present data show the possibility to select Ph- clones that (1) have a maintained capability of stroma adherence, (2) are mafosfamide resistant, (3) are derived from the CD34+ fraction, and (4) have high-replating potential.
Adult; Aged; Antigens; CD; CD34; Bone Marrow; Cell Adhesion; Cell Separation; Female; Fusion Proteins; bcr-abl; Granulocytes; Hematopoietic Stem Cells; Humans; In Situ Hybridization; Fluorescence; Leukemia; Myelogenous; Chronic; BCR-ABL Positive; Macrophages; Male; Middle Aged
6
info:eu-repo/semantics/article
262
Carlo Stella, C; Mangoni, L; Piovani, Giovanna; Garau, D; Almici, C; Rizzoli, V.
1 Contributo su Rivista::1.1 Articolo in rivista
none
1.1 Articolo in rivista
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11379/35143
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