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Tissue-Specific Stem Cells
Article first published online: 5 JAN 2009
Copyright © 2008 AlphaMed Press
Volume 27, Issue 1, pages 175–182, January 2009
How to Cite
Hayakawa, J., Hsieh, M. M., Uchida, N., Phang, O. and Tisdale, J. F. (2009), Busulfan Produces Efficient Human Cell Engraftment in NOD/LtSz-Scid IL2RγNull Mice. STEM CELLS, 27: 175–182. doi: 10.1634/stemcells.2008-0583
Author contributions: J.H.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; M.M.H.: conception and design, administrative support, manuscript writing, data analysis and interpretation; N.U.: data analysis and interpretation; O.P.: collection and assembly of data; J.F.T.: conception and design, financial support, administrative support, final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSExpress October 16, 2008.
- Issue published online: 5 JAN 2009
- Article first published online: 5 JAN 2009
- Manuscript Accepted: 28 SEP 2008
- Manuscript Received: 25 JUN 2008
- Cord blood;
- Peripheral blood stem cell transplantation;
- Severe combined immunodeficient mice;
- Xenograft transplant;
- Hematopoietic stem cells
Xenografting immunodeficient mice after low-dose irradiation has been used as a surrogate human hematopoietic stem cell (HSC) assay; however, irradiation requires strict and meticulous animal support and can produce significant mortality rates, limiting the usefulness of this model. In this work, we examined the use of parenteral busulfan as an alternative conditioning agent. Busulfan led to dose-dependent human HSC engraftment in NOD/LtSz-scid/IL2Rγnull mice, with marked improvement in survival rates. Terminally differentiated B and T lymphocytes made up most of the human CD45+ cells observed during the initial 5 weeks post-transplant when unselected cord blood (CB) products were infused, suggesting derivation from existing mature elements rather than HSCs. Beyond 5 weeks, CD34+-enriched products produced and sustained superior engraftment rates compared with unselected grafts (CB CD34+, 65.8% ± 5.35%, vs. whole CB, 4.27% ± 0.67%, at 24 weeks). CB CD34+ group achieved significantly higher levels of engraftment than mobilized CD34+-enriched peripheral blood (PB CD34+). At 8 weeks, all leukocyte subsets were detected, yet human red blood cells (RBCs) were not observed. Transfused human red cells persisted in the chimeric mice for up to 3 days; an accompanying rise in total bilirubin suggested hemolysis as a contributing factor to their clearance. Recipient mouse-derived human HSCs had the capacity to form erythroid colonies in vitro at various time points post-transplant in the presence of human transferrin (Tf). When human Tf was administered singly or in combination with anti-CD122 antibody and human cytokines, up to 0.1% human RBCs were detectable in the peripheral blood. This long evasive model should prove valuable for the study of human erythroid cells. STEM CELLS2009;27:175–182