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Sustained human hematopoiesis in immunodeficient mice by cotransplantation
of marrow stroma expressing human interleukin-3: analysis of gene
transduction of long-lived progenitors
JA Nolta, MB Hanley and DB Kohn
Division of Research Immunology/Bone Marrow Transplantation, Childrens
Hospital Los Angeles, CA 90027.
We have developed a novel cotransplantation system in which gene-
transduced human CD34+ progenitor cells are transplanted into
immunodeficient (bnx) mice together with primary human bone marrow (BM)
stromal cells engineered to produce human interleukin-3 (IL-3). The IL-
3-secreting stroma produced sustained circulating levels of human IL-3 for
at least 4 months in the mice. The IL-3-secreting stroma, but not control
stroma, supported human hematopoiesis from the cotransplanted human BM
CD34+ progenitors for up to 9 months, such that an average of 6% of the
hematopoietic cells removed from the mice were of human origin (human
CD45+). Human multilineage progenitors were readily detected as
colony-forming units from the mouse marrow over this time period.
Retroviral-mediated transfer of the neomycin phosphotransferase gene or a
human glucocerebrosidase cDNA into the human CD34+ progenitor cells was
performed in vitro before cotransplantation. Human multilineage progenitors
were recovered from the marrow of the mice 4 to 9 months later and were
shown to contain the transduced genes. Mature human blood cells marked by
vector DNA circulated in the murine peripheral blood throughout this time
period. This xenograft system will be useful in the study of gene
transduction of human hematopoietic stem cells, by tracing the development
of individually marked BM stem cells into mature blood cells of different
lineages.
Volume 83,
Issue 10,
pp. 3041-3051,
05/15/1994
Copyright © 1994 by The American Society of Hematology
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