Chemotaxis of primitive hematopoietic cells in response to stromal cell-derived factor-1.
Stromal cell-derived factor-1 (SDF-1) provides a potent chemotactic stimulus for CD34(+) hematopoietic cells. We cultured mobilized peripheral blood (PB) and umbilical cord blood (CB) for up to 5 weeks and examined the migratory activity of cobblestone area-forming cells (CAFCs) and long-term culture-initiating cells (LTC-ICs) in a transwell assay. In this system, SDF-1 or MS-5 marrow stromal cells placed in the lower chamber induced transmembrane and transendothelial migration by 2- and 5-week-old CAFCs and LTC-ICs in 3 hours. Transmigration was blocked by preincubation of input CD34(+) cells with antibody to CXCR4. Transendothelial migration of CB CAFCs and LTC-ICs was higher than that of PB. We expanded CD34(+) cells from CB in serum-free medium with thrombopoietin, flk-2 ligand, and c-kit ligand, with or without IL-3 and found that CAFCs cultured in the absence of IL-3 had a chemotactic response equivalent to noncultured cells, even after 5 weeks. However, addition of IL-3 to the culture reduced this response by 20-50%. These data indicate that SDF-1 induces chemotaxis of primitive hematopoietic cells signaling through CXCR4 and that the chemoattraction could be downmodulated by culture ex vivo.
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Laboratory of Developmental Hematopoiesis, Cell Biology Program, Sloan-Kettering Institute, New York, New York 10021, USA.
Rafii
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Hamada
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Moore
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D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000502:physiology; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D019718:Receptors, CXCR4 / Q000032:analysis / Q000502:physiology; D019089:Stem Cell Factor / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology
D018952:Antigens, CD34; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; C515256:Cxcl12 protein, mouse; D008565:Membrane Proteins; D019718:Receptors, CXCR4; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D013926:Thrombopoietin