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Blood, 1 November 2004, Vol. 104, No. 9, pp. 2919-2925.
Prepublished online as a Blood First Edition Paper on July 8, 2004; DOI 10.1182/blood-2004-03-0901.
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Submitted March 10, 2004
Accepted June 27, 2004
Characterisation of Acute Lymphoblastic Leukaemia progenitor cells
Charlotte V Cox, Roger S Evely, Anthony Oakhill, Derwood H Pamphilon, Nicholas J Goulden, and Allison Blair*
Bristol Institute for Transfusion Sciences, Bristol, United Kingdom
Bristol Haematology and Oncology Centre, Bristol, United Kingdom
Bristol Royal Hospital for Children, Bristol, United Kingdom
Bristol Royal Hospital for Children, Bristol, United Kingdom; Department of Pathology and Microbiology, University of Bristol, Bristol, United Kingdom
Bristol Institute for Transfusion Sciences, Bristol, United Kingdom; Department of Pathology and Microbiology, University of Bristol, Bristol, United Kingdom
* Corresponding author; email: allison.blair{at}nbs.nhs.uk.
Only a minority of acute lymphoblastic leukaemia (ALL) cells are thought to be capable of proliferating to maintain the leukaemic clone and these cells may be the most relevant to target with treatment regimens. We have developed a serum free suspension culture (SC) system that supported growth of B-ALL cells from 33 patients for up to 6 weeks. ALL cells from 28 cases (85%) were expanded in this system and growth was superior in SC than in long-term bone marrow culture. In order to characterise ALL progenitors, cells were sorted for expression of CD34 and CD10 or CD19 and the subfractions assayed in SC and in NOD/SCID mice. Cells capable of long-term proliferation in vitro and NOD/SCID repopulation were derived only from the CD34+/CD10- and CD34+/CD19- subfractions and these cells could engraft secondary recipients. The engrafted cells had the same immunophenotype and karyotype as was seen at diagnosis, suggesting they had differentiated in vivo. These results demonstrate that ALL cells capable of long-term proliferation in vitro and in vivo are CD34+/CD10-/CD19-. This suggests that cells with a more immature phenotype, rather than committed B-lymphoid cells, may be the targets for transformation in B-ALL.
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