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Blood, Vol. 111, Issue 4, 2444-2451, February 15, 2008
CD150– side population cells represent a functionally distinct population of long-term hematopoietic stem cells
Blood Weksberg et al.
111: 2444
Supplemental materials for: Weksberg et al
Files in this Data Supplement:
- Figure S1. SLAM Non-SP cells do not contain HSC activity (JPG, 89.2 KB)
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(A) SLAM cells were divided into SP and Non-SP fractions for transplant. (B) SPKLS cells were sorted as a control population. (C) 500 SLAM-SP, SLAM-Non-SP, and SPKLS donor cells (CD45.2) were transplanted into lethally irradiated mice (CD45.1) in the presence of 200,000 whole bone marrow competitor cells, and peripheral blood contribution (% CD45.2+) was measured at 4 and 17 weeks after transplant. No contribution to recipient hematopoiesis (<0.1%) was detected in recipients of SLAM-Non-SP cells (†).
- Figure S2. The addition of Sca-1 and c-Kit greatly improves the overlap between SLAM and SP cells (JPG, 56.2 KB)
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When CD150+48−41− cells are selected based on isotype controls, and the c-Kit+ Sca-1+ cells are then selected (middle panel), approximately 86% of these cells are in the SP.
- Figure S3. CD150+ and CD150− SPKLS HSCs are both capable of engrafting secondary recipients (JPG, 46.2 KB)
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At 24 weeks following primary transplantation, whole bone marrow from primary recipients was isolated and transplanted into new recipient mice (secondary recipients). Peripheral blood chimerism was measured at 4 and 8 weeks post-transplant. All three donor populations (SPKLS, CD150+ SPKLS, and CD150− SPKLS) were able to provide reconstitution in secondary transplants. A significant difference in secondary reconstitution was observed (with CD150− SPKLS showing diminished peripheral blood chimerism), however conclusions about qualitative differences cannot be assessed since the marrow used for secondary transplantation was not normalized to account for differences in bone marrow engraftment of purified HSCs in the primary cohorts (derived from experiments shown in Fig. 5).
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