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Blood, Vol. 106, Issue 5, 1574-1580, September 1, 2005
A novel stem-cell population in adult liver with potent hematopoietic-reconstitution activity
Blood Kotton et al.
106: 1574
Supplemental Data for: Kotton et al, Vol 106, Issue 5, 1574-1580
Files in this Data Supplement:
- Table S1. Summary of competitive blood repopulation assays (PDF, 85 KB) -
CD45.1+ peripheral blood chimerism resulting after transplantation of various cell populations in competition with 2 × 105 unfractionated CD45.2 bone marrow cells. Percentages represent average blood chimerism measured by fluorescence-activated cell sorting (FACS) 3 months after transplant (n ≥ 4 per group; SP = side population; PBMNC = peripheral blood mononuclear cells).
- Figure S1. Purification of CD45+ SP tip cells from adult liver (PDF, 69 KB) -
The gating algorithm and FACS plots demonstrate the approach for isolation of very rare cells from adult liver with potent hematopoietic reconstituting activity. After Ficoll gradient centrifugation and fluorescent staining (as detailed in “Materials and Methods”), the resulting cell suspension was analyzed by flow cytometry. First, a live cell gate was created excluding cell fragments (low forward scatter) or events that contain high propidium iodide (PI) fluorescence. Cells within this live cell gate were displayed on a Hoechst red/Hoechst blue dot plot, and side population (SP) cells were identified. “SP tip” and “SP shoulder” populations were identified by Hoechst and CD45 staining patterns. PI–/CD45+ SP tip cells were then purified by FACS-based cell sorting.
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