Prevalence and growth characteristics of malignant stem cells in B-lineage acute lymphoblastic leukemia

• Published in: Blood 1997-05, Vol.89 (10), p.3735-3744
• Main Authors: NISHIGAKI, H, ITO, C, MANABE, A, KUMAGAI, M.-A, COUSTAN-SMITH, E, YANISHEVSKI, Y, BEHM, F. G, RAIMONDI, S. C, PUI, C.-H, CAMPANA, D
• Format: Article
• Language: English
• Subjects: Adipose Tissue - cytology; Adipose Tissue - metabolism; Antigens, CD19 - analysis; Antigens, Neoplasm - analysis; Biological and medical sciences; Bone Marrow Cells; Burkitt Lymphoma - pathology; Cell Division; Child; Coculture Techniques; Connective Tissue - metabolism; Connective Tissue Cells; Culture Media, Serum-Free; Hematologic and hematopoietic diseases; HLA-DR Antigens - analysis; Humans; Immunophenotyping; Interleukin-7 - metabolism; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Medical sciences; Neoplastic Stem Cells - pathology; Stem Cell Factor - metabolism; Tumor Cells, Cultured; Tumor Stem Cell Assay
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v89.10.3735

We used a stroma-supported culture method to study the prevalence and growth characteristics of malignant stem cells in acute lymphoblastic leukemia (ALL). In 51 of 108 B-lineage ALL samples, bone marrow-derived stroma not only inhibited apoptosis of ALL cells but also supported their proliferation in serum-free medium. When single leukemic cells were placed in the stroma-coated wells of microtiter plates, the percentage of wells with leukemic cell growth after 2 to 5 months of culture ranged from 6% to 20% (median, 15%; 5 experiments). The immunophenotypes and genetic features of cells recovered from these cultures were identical to those noted before culture. All cells maintained their stroma dependency and self-renewal capacity. Leukemic clones derived from single cells contained approximately 10(3) to 10(6) cells after 1 month of culture; other clones became detectable only after prolonged culture. Cell growth in stroma-coated wells correlated with the number of initially seeded cells (1 or 10; r = .87). However, the observed percentages of positive wells seeded with 10 cells always exceeded values predicted from results with single-cell-initiated cultures (P < .003 by paired t-test), suggesting stimulation of leukemic cell growth by paracrine factors. In conclusion, the proportion of ALL cells with clonogenic potential may be considerably higher than previously thought.