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a Department of Cell Biology and Genetics, Erasmus University, Rotterdam, Netherlands;
b Third Department of Internal Medicine, Technical University of Munich, Germany;
c Max Planck Institute of Molecular Genetics, Berlin, Germany
Key Words. Hematopoiesis • Hematopoietic stem cells • AGM • Transwell • Microenvironment
Correspondence: Robert A.J. Oostendorp, Ph.D., III Medizinische Klinik und Poliklinik, Klinikum Rechts der Isar, Technische Universität München, Ismaningerstrasse 22, 81675 München, Germany. Telephone: 49-89-4140-2362; Fax: 49-89-4140-4826, e-mail: oostendorp{at}lrz.tum.de
We recently established that two midgestation-derived stromal clones—UG26-1B6, urogenital ridge–derived, and EL08-1D2, embryonic liver–derived—support the maintenance of murine adult bone marrow and human cord blood hematopoietic repopulating stem cells (HSCs). In this study, we investigate whether direct HSC-stroma contact is required for this stem cell maintenance. Adult bone marrow ckit+ Ly-6C– side population (K6-SP) cells and stromal cells were cocultured under contact or noncontact conditions. These experiments showed that HSCs were maintained for at least 4 weeks in culture and that direct contact between HSCs and stromal cells was not required. To find out which factors might be involved in HSC maintenance, we compared the gene expression profile of EL08-1D2 and UG26-1B6 with four HSC-nonsupportive clones. We found that EL08-1D2 and UG26-1B6 both expressed 21 genes at a higher level, including the putative secreted factors fibroblast growth factor-7, insulin-like growth factor-binding proteins 3 and 4, pleiotrophin, pentaxin-related, and thrombospondin 2, whereas 11 genes, including GPX-3 and HSP27, were expressed at a lower level. In summary, we show for the first time long-term maintenance of adult bone marrow HSCs in stroma noncontact cultures and identify some secreted molecules that may be involved in this support.
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