Prevalence and growth characteristics of malignant stem cells in B-lineage acute lymphoblastic leukemia.
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.
Nishigaki
H
H
Department of Hematology-Oncology, St Jude Children's Research Hospital, University of Tennessee College of Medicine, Memphis 38105, USA.
Ito
C
C
Manabe
A
A
Kumagai
M
M
Coustan-Smith
E
E
Yanishevski
Y
Y
Behm
F G
FG
Raimondi
S C
SC
Pui
C H
CH
Campana
D
D
D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism; D018941:Antigens, CD19 / Q000032:analysis; D000951:Antigens, Neoplasm / Q000032:analysis; D001854:Bone Marrow Cells; D002051:Burkitt Lymphoma / Q000473:pathology*; D002455:Cell Division; D002648:Child; D018920:Coculture Techniques; D003238:Connective Tissue / Q000378:metabolism; D003239:Connective Tissue Cells; D016895:Culture Media, Serum-Free; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D016130:Immunophenotyping; D015851:Interleukin-7 / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000473:pathology*; D019089:Stem Cell Factor / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
D018941:Antigens, CD19; D000951:Antigens, Neoplasm; D016895:Culture Media, Serum-Free; D006684:HLA-DR Antigens; D015851:Interleukin-7; D019089:Stem Cell Factor