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Engraftment and Retroviral Marking of CD34+ and
CD34+CD38 Human Hematopoietic Progenitors
Assessed in Immune-Deficient Mice
Mo A. Dao,
Ami J. Shah,
Gay M. Crooks, and
Jan A. Nolta
From the Division of Research Immunology/Bone Marrow Transplantation,
Childrens Hospital Los Angeles, Los Angeles, CA, and University of
Southern California School of Medicine, Department of Pediatrics, Los
Angeles, CA.
Retroviral-mediated transduction of human hematopoietic stem cells
to provide a lifelong supply of corrected progeny remains the most
daunting challenge to the success of human gene therapy. The paucity of
assays to examine transduction of pluripotent human stem cells hampers
progress toward this goal. By using the beige/nude/xid (bnx)/hu immune-deficient mouse xenograft system, we
compared the transduction and engraftment of human CD34+
progenitors with that of a more primitive and quiescent subpopulation, the CD34+CD38 cells. Comparable extents of
human engraftment and lineage development were obtained from 5 × 105 CD34+ cells and 2,000 CD34+CD38 cells. Retroviral marking of
long-lived progenitors from the CD34+ populations was
readily accomplished, but CD34+CD38 cells
capable of reconstituting bnx mice were resistant to transduction. Extending the duration of transduction from 3 to 7 days
resulted in low levels of transduction of
CD34+CD38 cells. Flt3 ligand was required
during the 7-day ex vivo culture to maintain the ability of the cells
to sustain long-term engraftment and hematopoiesis in the mice.
Blood, Vol. 91 No. 4 (February 15), 1998:
pp. 1243-1255
© 1998 by The American Society of Hematology.
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