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Engraftment of Cultured Human Hematopoietic Cells in Sheep
Yoshifumi Shimizu,
Makio Ogawa,
Masao Kobayashi,
Graca Almeida-Porada, and
Esmail D. Zanjani
From the Department of Medicine, Medical University of South
Carolina, Charleston, SC; and Department of Veterans
Affairs Medical Centers, Charleston, SC and Reno, NV.
In an effort to expand human hematopoietic progenitors and stem
cells in vitro, we cultured human
CD34+c-kitlow bone marrow cells in suspension
in the presence of KIT ligand, FLK2/FLT3 ligand, interleukin-6 (IL-6),
and erythropoietin with or without IL-3 and tested their engrafting
capabilities by injecting them into sheep fetuses. As markers for
engraftment, we analyzed CD45+ cells and karyotypes of
the colonies grown in methylcellulose culture. In three separate
experiments, day-60 engraftment in the bone marrow was seen with both
fresh cells and cells cultured in the presence or absence of IL-3. When
fetuses were allowed to be born and analyzed for CD45+
cells, no long-term engraftment was seen with cultured cells. We then
pooled the CD45+ cells of the fetal samples and
transplanted them into secondary recipient fetuses. Day-60 engraftment
in the secondary recipients was again noted when transplantation in the
primary recipients was initiated with fresh cells. There were 3 cases
in which cultured cells showed signs of engraftment in the secondary
recipients, but the remaining 24 cases showed no signs of engraftment.
These data documented that suspension culture for 2 weeks of enriched adult human bone marrow cells can maintain short-term (2 months) engrafting cells, but may not maintain longer term engrafting cells.
This sheep/human xenograft model may serve as an excellent method for
the evaluation of the engraftment potential of in vitro-expanded cells.
Blood, Vol. 91 No. 10 (May 15), 1998:
pp. 3688-3692
© 1998 by The American Society of Hematology.
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