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Blood, 15 January 2005, Vol. 105, No. 2, pp. 576-583.
Prepublished online as a Blood First Edition Paper on September 2, 2004; DOI 10.1182/blood-2004-04-1467.
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Submitted April 20, 2004
Accepted August 23, 2004
Soluble factors elaborated by human brain endothelial cells induce the concomitant expansion of purified human BM CD34+CD38- cells and SCID repopulating cells
John P Chute*, Garrett G Muramoto, Jennifer Fung, and Carol Oxford
Stem Cell Biology Laboratory, Large Scale Biology Corporation, Vacaville, CA, USA; Division of Hematology/Oncology, UC Davis Cancer Center, Sacramento, CA, USA
Stem Cell Biology Laboratory, Large Scale Biology Corporation, Vacaville, CA, USA
The Jackson Laboratories, Sacramento, CA, USA
Department of Pathology, UC Davis, Davis, CA, USA
* Corresponding author; email: john.chute{at}duke.edu.
The CD34+CD38- phenotype identifies a population in the bone marrow that is enriched in the steady state for hematopoietic stem cells (HSC). Following ex vivo culture of CD34+ cells, HSC content is difficult to measure since committed CD34+CD38+ progenitors down regulate CD38 surface expression during culture. In this study, we sought to define the phenotype of human HSC following ex vivo culture under conditions which support the expansion of primitive SCID-repopulating cells (SRC). Contact co-culture of FACS-sorted BM CD34+CD38- cells with human brain endothelial cells (HUBEC) supported a 4.4-fold increase in CD34+CD38- cells with a concordant 3.6-fold increase in SRC over 7 days. Non-contact HUBEC cultures and the addition of thrombopoietin, SCF, and Flt-3 ligand supported further increases in CD34+CD38- cells (6.4-fold and 13.1-fold) which correlated with significant increases in SRC activity. Moreover, cell sorting studies performed on HUBEC-cultured populations demonstrated that SRC were significantly enriched within the CD34+CD38- subset as compared to the CD34-CD38- population post-culture. These results indicate that human HSC can be identified and characterized by phenotype following expansion culture. These studies also demonstrate that HUBEC-elaborated soluble factors mediate a unique and potent expansion of human HSC.

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