| Autor: |
Cerny, Jan, Dooner, Mark, McAuliffe, Christina, Habibian, Houri, Stencil, Kimberly, Berrios, Virla, Reilly, Judy, Carlson, Jane, Cerny, Anna Maria, d'Hondt, Lionel, Benoit, Brian, Lambert, Jean-Francois, Colvin, Gerald, Nilsson, Susan, Becker, Pamela, Quesenberry, Peter |
| Zdroj: |
Journal of Hematotherapy & Stem Cell Research (now called Stem Cells and Development); December 01, 2002, Vol. 11 Issue: 6 p913-922, 10p |
| Abstrakt: |
This study was designed to establish a direct homing assay using purified lineage-negative Sca-1-positive (Lin- Sca+) murine bone marrow cells and to evaluate the effects of cytokines on homing. C57BL/6 Lin- Sca+ marrow stem cells were labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then injected by tail vein into untreated C57BL/6 mice. Marrow was harvested at various times after cell infusion and analyzed on a high-speed MoFlo cell sorter for fluorescent positive events, using a large event analysis, with at least 16 million total events analyzed. We have shown that homing of Lin- Sca+ cells plateaus by 1 h, and at 3 h post-infusion is linear between 50,000 and 1,000,000 infused cells. This forms a base for a homing assay in which 250,000 CFDA-SE labeled Lin- Sca+ marrow cells are infused and then recovered from marrow 3 h later, followed by a large-event fluorescence-activated cell sorting (FACS) analysis. We found that 7.45-9.32% of infused cells homed and that homing of stem cells cultured for 48 h in interleukin-3 (IL-3), IL-6, IL-11, and steel factor cultured cells was defective when compared to noncultured cells. Exposure of marrow stem cells to IL-3, IL-6, IL-11, and steel factor induces a stem cell homing defect, which probably underlies the engraftment defect previously characterized under these conditions. |
| Databáze: |
Supplemental Index |
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