Homing of purified murine lymphohematopoietic stem cells: a cytokine-induced defect
| Autor: | Lionel D'Hondt, Peter J. Quesenberry, J. Reilly, Kimberly Stencil, Gerald A. Colvin, Jan Cerny, Pamela S. Becker, Jane E. Carlson, Houri Habibian, Mark S. Dooner, Anna M. Cerny, Jean Francois Lambert, Christina I. McAuliffe, Brian O. Benoit, Susan K. Nilsson, Virla M. Berrios |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Male
medicine.medical_treatment Immunology Cell Cell Culture Techniques Bone Marrow Cells Thymus Gland Biology Mice Cell Movement medicine Animals Bone Marrow Transplantation Stem Cell Factor Interleukin-6 Graft Survival Tail vein Hematology Cell sorting Hematopoietic Stem Cells Interleukin-11 Molecular biology Antigens Differentiation Mice Inbred C57BL Kinetics medicine.anatomical_structure Cytokine Cytokines Interleukin-3 Bone marrow Stem cell Event analysis Spleen Homing (hematopoietic) |
| Zdroj: | Journal of hematotherapystem cell research. 11(6) |
| ISSN: | 1525-8165 |
| Popis: | 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: | OpenAIRE |
| Externí odkaz: |
|